Formulations And Methods For Treating Photosynthetic Organisms And Enhancing Qualities And Quantities Of Yields With Glycan Composite Formulations

ABSTRACT

Glycan Composites and methods for rendering glycan composites for the treatment of photosynthetic organisms, including the steps of formulating branched glycan deglycosylates into coordination complex compositions resulting in water-borne availability; stability during storage; applying a suitable volume of the resulting mixture to one or more photosynthetic organisms; delivery to photosynthetic organisms; metabolically based growth of crops; enhanced qualities and increased quantities of crops; and systems and compositions for the same.

CROSS-REFERENCE

This application is a divisional of U.S. patent application Ser. No.15/492,110 filed Apr. 20, 2017, which claims priority of ProvisionalApplication Ser. No. 62/329,226 filed Apr. 29, 2016, the disclosures ofwhich are incorporated herein by reference.

FIELD

Embodiments disclosed herein relate to methods for treatingphotosynthetic organisms, such as fields of photosynthetic organisms,including farm crops, and/or enhancing their growth with formulationsthat comprise compositions of glycan composites comprising branchedglycan deglycosylate components with coordination complex components.Moreover, in liquid compositions of matter, the foregoing formulationsmay include one or more preservatives, rendering the formulationssuitable to ship and store. Embodiments disclosed herein include systemsfor delivery of glycan composites into photosynthetic organisms.

BACKGROUND

Embodiments disclosed herein relate to the application to photosyntheticorganisms of glycan composite formulations comprised of one or morebranched glycan deglycosylates and transition metal²⁺ coordinationcomplexes.

The health of photosynthetic organisms in agricultural crops isdependent on their biological manufacture of photosynthates, especially,sugars; and compositions of the present embodiments enhance theavailability of these photosynthates to promote crop health and growth.Recent major advancements for crop improvements include water-solubleglucosides as described in the patent literature. While these glucosideshave proven effective at foliar rates ranging in the application ofkilograms per hectare, there is a need for formulations that are morepotent and effective in the range of grams per hectare as shown in theembodiments herein. The branched glycan deglycosylates of the presentembodiments are of higher order potencies than conventional compositionswhile transition metal²⁺ coordination complex components of the glycancomposites further improve activity. In addition, by treatment of sapnectar, glycan composites improve photosynthate flux capacity of a crop.Further still, methods and compositions of glycan composites may becustomized for improvement of qualities and/or quantities of crops whilesustaining potency of the glycan composite.

SUMMARY

It is an object of embodiments disclosed herein to provide methods andformulations for treating photosynthetic organisms and enhancingqualities and quantities of crop yields. The formulations comprise oneor more branched glycan deglycosylates combined with transition metal²⁺coordination complexes, thereby forming a glycan composite. Embodimentsprovide methods and compositions for the formulation of glycancomposites.

It is a further object of embodiments disclosed herein to provide liquidformulations comprising one or more formulations of glycan compositeswith preservatives for retention of high potency.

It is a further object of embodiments disclosed herein to providemethods and formulations for treating photosynthetic organisms such asplants, and for enhancing photosynthetic organismal growth, such as inplant crops, by applying a glycan composite formulation.

It is another object of embodiments disclosed herein to provide glycancomposite formulations comprising metals²⁺ components of transitionmetal coordination complexes and providing anionic components oftransition metal²⁺ coordination complexes.

It is a further object of embodiments disclosed herein to provide glycancomposite formulations comprising soluble transition metals²⁺ preferablyin coordination complexes.

It is a further object of embodiments disclosed herein to provide glycancomposite formulations comprising one or more anionic components of thetransition metal²⁺ coordination complexes.

It is a further object of embodiments disclosed herein to provide glycancomposite formulations comprising one or more anionic components of thetransition metal²⁺ coordination complexes, wherein the anionic componentis selected from polydentate sequestering agents.

It is a further object of embodiments disclosed herein to provide amethod for formulating a glycan composite composition that comprises oneor more preservative components that maintains active liquid solutionsthrough periods of shipping and storage.

It is a further object to provide methods for treatment ofphotosynthetic organisms, particularly plants, and growth formulationsfor photosynthetic organisms comprising glycan composite formulationsrendered as liquid and/or dry packages that are available andpenetrative consistent with facilitation of transcuticular,transepidermal and/or transmembrane transport; seed, foliar and rootuptake of solutes; germination; and/or that maintain growth in low lightintensity environments.

It is a further object to provide treatment and growth formulations forphotosynthetic organisms and to their parts; and particularly applied toagricultural crops.

It is a further object to provide a plant seed treatment formulationcomprising glycan composite compositions and methods for management ofgrowth in green plants, particularly where applied to seeds to hastengermination and growth.

It is a further object to provide floral and/or fruit treatmentformulations comprising glycan composite compositions for management ofquality growth in photosynthetic organisms; particularly where appliedto roots, foliage, flowers and/or fruit while attached to the plantbefore harvest; to detached flowers and/or fruit after harvest; and/orto improve flavor qualities.

It is another object to provide glycan composite compositions of matter.

It is another object to provide glycan composites comprised of branchedglycan deglycosylates and soluble transition metals²⁺.

It is another object to provide glycan composites, comprised of branchedglycan deglycosylates obtained from natural products.

It is another object to provide a glycan composite and/or its componentsas novel plant growth regulators selected to increase crop quantity andquality.

It is another object to provide novel glycan composite plant growthregulators for the management of respiration in photosyntheticorganisms.

It is yet another object to provide a glycan composite environment inthe presence of photosynthetic organisms under cultivation conducive ofrespiration in low light intensity shaded to dark conditions.

It is another object to provide an environment of respiratoryacceleration in the presence of photosynthetic organisms undercultivation by treatments with glycan composites. In some embodiments,roots at any time of day or night, and/or shoots in the dark of night,are exposed to respiration accelerators that elevate ambient oxygen (O₂)either through >25% O₂ gas treatment of a photosynthetic organism, or byapplication of O₂-generators. Furthermore, another object is to providean environment conducive to respiration in the presence of thephotosynthetic organisms under cultivation by treatments with glycancomposites; wherein photosynthetic organisms under cultivation bytreatments with glycan composites may be exposed to respirationaccelerators.

It is yet another object to provide an environment conducive to theaccumulation of photosynthates in the presence of photosyntheticorganisms under cultivation by treatments with glycan composites.Photosynthetic organisms under cultivation by treatments with glycancomposites may be exposed to respiration decelerators conducive to theaccumulation of photosynthates.

It is another object to provide glycan composites and/or theircomponents as natural product biostimulants selected to benefit nutrientuse efficiency, improve tolerance to abiotic stress, and/or increasecrop quantity and quality.

It is another object to provide glycan composite treatments forphotosynthetic growth, comprising branched glycan deglycosylates andtransition metal²⁺ coordination complexes. In certain embodiments, suchas for convenient utilization in the field, the complex is renderedconvenient to apply to photosynthetic organisms and readily safe forphotosynthetic organisms by formulation in liquid and/or dry packagesand with the option for admixture of crop treatments.

It is a further object to provide treatment methods and compositions forphotosynthetic organisms comprising exogenous glycan compositecompositions for control of the endogenous deconjugation ofphotosynthates that advance the quality and quantity of commercialharvests.

It is a further object to provide a treatment and growth formulation forphotosynthetic organisms comprising exogenous glycan compositecompositions for treatment of endogenous photosynthates that enhance theflavor qualities of commercial yields.

It is a further object to provide a treatment and growth formulation forphotosynthetic organisms comprising glycan composite compositions thatare preserved for storage with one or more phytobland preservatives.

It is a further object to provide a treatment and growth formulation forphotosynthetic organisms comprising glycan composite systems for theimprovement of crop growth in water culture.

It is a further object to provide a treatment and growth formulation forphotosynthetic organisms comprising glycan composites as synergisticsystems for improving crop aesthetic quality by reduction of theincidence of sun scorch.

It is a further object to provide a method for treatment ofphotosynthetic organisms that fortifies the nutritional qualities of sapnectar of flowering plant fields comprising applying glycan compositesto flowering food plants for the vigorous health benefit of pollinatorsand grazers.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more methyl-α-D-Mannopyranosyl (Man)₁₋₃ componentsand one or more citrate-Ca²⁺-Mn²⁺-coordination complex components andoptionally with one or more D-block transition metals²⁺; and/or furtherformulated with one or more optional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more methyl-α-D-Man₁₋₃ components and one or moremalate-Ca²⁺-Mn²⁺-coordination complex components and optionally with oneor more D-block transition metals²⁺; and/or formulated with one or moreoptional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more methyl-α-D-Man₁₋₃ components and one or moreglutarate-Ca²⁺-Mn²⁺-coordination complex components; and optionally withone or more D-block transition metals²⁺; and/or further with one or moreoptional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more methyl-α-D-Man₁₋₃ components and one or moresuccinate-Ca²⁺-Mn²⁺-coordination complex components and optionally withone or more D-block transition metals²⁺; and/or further with one or moreoptional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more Man₁₋₃N-linked-glycan components; one or moreCa²⁺-Mn²⁺-citrate coordination complex components; and optionally withone or more D-block transition metals²⁺; and/or further with one or moreoptional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more Man₁₋₃N-linked-glycan components; one or moreCa²⁺-Mn²⁺-coordination complex components; optionally with one or moreD-block transition metals²⁺; and/or further with one or more optionalpreservatives

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more methyl-α-D-Glc₁ components; one or more citrateCa²⁺-Mn²⁺-coordination complex components; optionally with one or moreD-block transition metals²⁺; and/or further with one or more optionalpreservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more deglycosylated glycan components;Ca²⁺-Mn²⁺-coordination complex components; optionally with one or moreD-block transition metals²⁺; and/or further with one or more optionalpreservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more deglycosylated O-linked-glycan components frombotanical gums; Ca²⁺-Mn²⁺-coordination complex components; optionallywith one or more D-block transition metals²⁺; and/or further with one ormore optional preservatives.

In certain embodiments, a glycan composite is formulated as a complexcomprised of one or more deglycosylated N-linked-glycan components frominvertase; Ca²⁺-Mn²⁺-coordination complex components; optionally withone or more D-block transition metals²⁺; and/or further with one or moreoptional preservatives.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic representation of the flow of processes from topto bottom resulting in methods and compositions for the treatment ofphotosynthetic organisms with the glycan composite, in accordance withcertain embodiments. In the example of FIG. 1, the application of aglycan composite exposes a plant cell to a solution that enhancesqualities and quantities of crops. The glycan composite is transportedinto the cell; metabolized; and energy is transferred. This metabolicpathway, defined by the glycan composite routing photosynthates out ofstorage and into respiration of O₂→CO₂, led to heightened plantproductivity for enhanced quality and yield.

FIG. 2 exhibits drawings of exemplary branched glycan deglycosylatessuitable for formulation in glycan composites. The core structure in thetop left corner corresponds to a trimannopyranosyl-N-glycan whilst otherstructures display higher orders of branching. The invertase corestructures shown are suitable for selection of one or more branchedglycan deglycosylates with preferred terminal ligands of the glycancomposite. Exemplary high mannopyranosyl N-glycans with terminalmannopyranosyl ligands show progressions to higher orders of branchingfrom top to bottom of the page. The top left Man₃GlcNAc₂ structurecorresponds to “Ethan” in Examples 1, 6, 9, 16, and 17. Abbreviations:Glc—Glucopyranosyl; Man—Mannopyranosyl; NAc—N-Acetyl.

DETAILED DESCRIPTION

Unless otherwise defined, all technical and scientific terms employedherein have their conventional meaning in the art. As used herein, thefollowing terms have the meanings ascribed.

“M” refers to molar concentration, “μM” refers to microMolar, and “mM”refers to milliMolar concentration.

“kD” refers to kiloDalton

“PGR” refers to a plant growth regulator.

“Percent” or “%” is percent by weight unless otherwise indicated.

“Ppb” refers to parts per billion by weight.

“Ppm” refers to parts per million by weight.

“Ppt” refers to parts per thousand by weight.

Nomenclature—Botanical names generally are given as common names thatare referable to scientific nomenclature through abundant scholarlyresources.

Statistics—Comparison of means by T-test (two-tailed) show significanceat ρ≤0.05.

“Enhance growth” or “enhancing growth” refers to promoting, increasingor improving the rate of growth of a photosynthetic organism such as aplant; and/or increasing or promoting an increase in the size and/oryield; and/or enhancing the quality of the photosynthetic organism orits parts; regulating the flow of photosynthates; enhancing the flow ofphotosynthates to respiration; enhancing aesthetics; increasinghydrostatic pressure; improving fragrance; accumulating photosynthateswithin the photosynthetic organism; and/or improving the flavor of thephotosynthetic organism, in particular, Brix (a measure of sugarcontent), of its seed, fruit, flower, nectar, root, stem, or its parts.

“Photosynthetic Organism” refers to life forms that synthesizephotosynthates including C₃, C₄, and CAM plants; and photosyntheticEukaryotes including, but not necessarily limited to, those of thefollowing preferred supergroups: Archaeplastida such as Plantae,Chlorophyta and Rhodophyta; and Chromoveolata such as Phaeophyta.Photosynthetic organisms may also refer to botanicals; turf andornamentals; crops, including food, fodder, fiber, feed, andagricultural crops; and harvests thereof; and plants, both higher andlower plants, and plant-like organisms. The systems, methods andformulations may be advantageously used with any species ofphotosynthetic life.

The compositions may be applied to virtually any variety of livephotosynthetic organisms. Photosynthetic organisms which may benefitinclude, but are not limited to, all Plantae particularly those in allcrop groups recognized by the United States Environmental ProtectionAgency (2012: 40 CFR 180.41) as for example such as the following:alfalfa, allspice, amaranth, angelica, anise, annatto, arugula, backciao, balm, barley, basil, bean, beet, borage, breadfruit, broccoli,Brussels sprouts, burdock, burnet, cabbage, cantaloupe, caper, caraway,cardamom, cardoon, carrot, cassava, castor, cauliflower, cavalo,broccolo, celeriac, celery, celtuce, cereals, chard, chayote, chervil,chickpea, chicory, chive, cilantro, cinnamon, clove, clover, coffee,collards, coriander, corn, cotton, cranberry, cress, cucumber, cumin,curry, daikon, daylily, dill, endive, euphorbia, eggplant, fennel,fenugreek, flax, forage, fritillaria, gherkin, gourd, grape, grain,garlic, guar, hay, hemp, horehound, hosta, hyssop, jackbean, jicama,jojoba, kale, kohlrabi, kudzu, kurrat, lablab bean, lavender, leafygreens, leek, legume, lemongrass, lentil, lespedeza, lettuce, lupin,mace, marjoram, melon, millet, mint, mizuna, Momordica, muskmelon,mustard, nasturtium, nutmeg, oat, onion, orach, parsley, parsnip,pasture, pea, peanut, pepper, peppermint, perilla, popcorn, potato,poppy, pumpkin, purslane, radicchio, radish, rape greens, rhubarb, rice,rosemary, rutabaga, rye, safflower, saffron, sage, sainfoin, salsify,skirret, sesame, shallot, sorghum, soybean, spinach, squash, stevia,strawberry, sunflower, sweet bay, sweet potato, sugar beet, sugar cane,Swiss chard, swordbean, tanier, taro, tarragon, tea, teosinte, thyme,tobacco, tomato, trefoil, triticale, turmeric, turnip, vanilla,vernonia, vetch, watermelon, wheat, wild rice, wintergreen, woodruff,wormwood, yam, zucchini and the like; fruit-bearing plants, such as,almond, apple, apricot, avocado, azarole, banana, beech nut, blackberry,blueberry, Brazil nut, breadfruit, butternut, cashew, cherry, chestnut,chinquapin, citrus, cocoa, cocona, coffee, currant, dragonfruit,elderberry, fig, filbert, goji, gooseberry, grapefruit, guava, hickorynut, huckleberry, kiwifruit, kumquat, lemon, lime, loganberry, loquat,macadamia nut, mango, mangosteen, martynia, mayhaw, naranjilla,nectarine, nopales, nut, okra, olive, orange, papaya, passion fruit,peach, pear, pecan, pepper, pistachio, plum, plumcot, prune, pummelo,quince, raspberry, roselle, tangelo, tangerine, tangor, tejocote,tomatillo, uniq fruit, walnut, spices, and the like; florals andornamentals, such as achillea, adenium, agave, ageratum, aloe, alyssum,anemone, aquilegia, aster, azalea, begonia, bird-of-paradise, bleedingheart, borage, bromeliad, bougainvillea, buddlea, cactus, calendula,camellia, campanula, carex, carnation, celosia, chrysanthemum, clematis,cleome, coleus, cosmos, crocus, croton, cyclamen, dahlia, daffodil,daisy, dandelion, day lily, delphinium, dianthus, dietes, digitalis,dipladenia, dock, dusty miller, euonymus, forget-me-not, fremontia,fuchsia, gardenia, gazania, geranium, gerbera, gesneriad, gladiolus,hibiscus, hydrangea, impatiens, jasmine, lily, lilac, lisianthus,lobelia, marigold, mesembryanthemum, mimulus, myosotis, narcissus, NewGuinea Impatiens, nymphaea, oenothera, oleander, orchid, ornamentals,oxalis, pansy, penstemon, peony, petunia, plumeria, poinsettia,polemonium, polygonum, poppy, portulaca, primula, ranunculus,rhododendron, rose, salvia, senecio, shooting star, snapdragon, solanum,solidago, stock, ti, torenia, tulip, verbena, vinca, viola, violet,yucca, zinnia, and the like; indoor garden and houseplants, such asAfrican violet, Chinese evergreen, succulents, dieffenbachia, dracaena,ficus, hosta, peace lily, philodendron, pothos, rubber tree,sansevieria, chlorophytum, and the like; trees, such as Abies, Aspen,birch, cedar, Cinnamomum, Cornus, cycad, cypress, Dawn Redwood, elm,ficus, fir, ginkgo, gymnosperm, hardwood trees, Indian Rosewood,jacaranda, juniper, Laurel, legume, Liriodendron, magnolia, mahogany,maple, oak, palm, Picea, Pinus, Pittosporum, Plantago, poplar, redwood,rosewood, saguaro, Salix, sycamore, Taxus, teak, willow, yew, Christmastree, sources of lumber, sources of paper, and the like; grasses, suchas turf, sod, bluegrass, bent grass, Bermuda grass, bromegrass,calamogrostis, carex, creeping bent, elymus, fescue, festuca,helictotrichon, imperata, miscanthus, molina, panicum, paspalum,pennisetum, phalaris, poa, grass seeds, and the like; dwarfs; grafts;cuttings; hybrids; and the like. In addition to the aforementionedcrops, the formulations are also suitable for application tophotosynthetic organismal sources of secondary metabolites such asswitchgrass, Jatropha, euphorbia, nicotiana, lichen, kelp, diatom,cyanobacteria, bacteria, dunaliella, nannochloropsis, chlorella,haematococcus, eucheuma; bryophytes such as moss and fern; and the like.This list is intended to be exemplary and is not intended to beexclusive. Other photosynthetic organisms that may benefit byapplication of the compositions and methods of the present embodimentswill be readily determined by those skilled in the art. The methods andformulations disclosed herein may be used to enhance growth in juvenileand mature photosynthetic organisms, as well as cuttings, tissues,seeds, meristems, callus, cells, and micropropagation. Seed priming andcoatings prior to sowing may be applied in the range of 10-1000 μg ofglycan composites per seed, preferably in the range of 20-300 μg/seed.

Alternatively, seeds, corms, bulbs, stolons, and cuttings, may betreated in-furrow, simultaneously with sowing. Generally, the anatomicallocation to which the composition of the method is applied should have asurface area large enough to enable the photosynthetic organism toabsorb the composition. For example, it is desirable to include thesprouted cotyledon (i.e., the “seed leaf”), potato stolon, bulb, corm,or other substantial surfaces that facilitate absorption, such as trueleaves and roots. Fruit bearing plants may be treated before and afterthe onset of bud, fruit and seed formation. For plants such as annuals,perennials, trees, orchids, gesneriads, and cacti in which stems, rootsand/or trunks may be treated, application methods include treatment ofshoots with sprays and/or treatment of shoots and roots by sprench ordip application or by separate root and shoot applications. Commercialaqua- and mariculture crops such as spirulina, aonori, laver, kombu,macrocystis, nori and wakame, may be misted, sprayed, brushed or dippedin sterile aqueous freshwater or seawater solutions of 10 ppb-3% glycancomposites, allowing 15-90 minutes to absorb.

The Glycan Composite

The methods and formulations in accordance with certain embodimentsdisclosed herein are designed, for example, to treat any of theaforementioned photosynthetic organisms such as plants, and to enhancequality, increase growth and/or improve the quality and quantity ofharvested yields. This can be achieved by applying glycan compositeformulations comprised of the following: one or more branched glycandeglycosylates with certain transition metal²⁺coordination complexes.The formulations can be applied in a dry or liquid form directly tophotosynthetic organisms. In certain embodiments, liquid formulationsadditionally may include a preservative for prevention of spoilageduring shipping and storage periods. The methods disclosed herein makeglycan composites readily available for uptake by photosyntheticorganisms.

The Branched Glycan Deglycosylate Component

Certain embodiments disclosed herein provide branched glycandeglycosylates that are components of the glycan composite. Hereinafter,this branched glycan deglycosylate component will be referred to as the“glycan” or “deglycosylate” component of the glycan composite.

Glycans are rather expensive when chemically synthesized and, if not forcertain of the embodiments of the invention, agricultural applicationswould not be economically justifiable.

Fortunately, embodiments disclosed herein provide a number ofcost-effective products by means of deglycosylation of certaininexpensive macromolecules, and these products make suitable glycancomponents of the glycan composite. Thus, certain branched glycandeglycosylates render economically feasible farm crop treatments.Suitable glycans may originate by cleavage, i.e., deglycosylation ofglycan subunits from their parent macromolecule. Generally,macromolecules greater than 1000 s kD are chemical structures too largefor treatment and uptake by a photosynthetic organism; therefore,deglycosylates less than 10 s kD are preferred branched glycandeglycosylate components. Deglycosylates are typically frommacromolecules such as proteins, glycoproteins,N-linked-glycan-macromolecules and/or O-linked-glycan-macromolecules.They may be products of hydrolysis or other processes known to the art,resulting from actions of acids, bases, enzymes and/or microbes breakingbonds. Biosynthesis of branched glycan components by a plant or yeastmay be cost effective as compared to products of chemical synthesis andpurification, the expense of which has proven prohibitive. For example,purchase of pure high mannan branched N-linked glycans may cost $1000s/gram; whereas, suitable high mannan branched N-linked glycans,deglycosylated from proteins per embodiments disclosed herein, may costpennies/gram.

Botanical sources of suitable glycan subunits include the following:Cyanaposis tetragonalobus and Cyanaposis psoraloides, guar gums, GalMan₂; Caesalpinia spinosa, tara gums, GalMan₃ ; Ceratonia siliqua, locustbean gums, GalMan₁₋₈ ; Amorphophallus konjac, konjac gums, Glc₂Man₂ ;Canavalia ensiformis Jack Bean, N-linked glycans; Ivory nut, Man_(n);carob; coffee bean; fenugreek; barley; palms, lilies, irises, andlegumes, endosperm tissues, Man_(n); soft wood and bark of varioustrees; birch; gymnosperms; Norway spruce; and Chlorophyta such asDasycladales, Characeae, Codium fragile, Caulerpa and Acetabulariaacetabulum Mannan Weed. Furthermore, branched mannan derivativestructures such as exhibited in FIG. 2, may be found in fungi, such as,Hansenula holstii, Rhodotorula acheniorum; in glycoproteins, such as,concanavalins and enzymes; and preferably in invertases. Other naturalsources include microbes; bacteria; mushrooms; animals, such as,arthropods, crustaceans, shellfish, fish, krill, and insects; and waste,such as guano, offal, blood, marrow, liver, animal organ, bark, sawdust,wood, bone, exoskeleton, ferment, bycatch, and manure.

The aforementioned gums, proteins and other macromolecules may undergodeglycosylation by commercial processes known in the art. For example,some branched glycan macromolecules may be microbially digested undertightly controlled fermentation and others may be subjected to variousother enzymatic digestion processes known in the art; and whereby,branched glycan macromolecules may be partially hydrolyzed bycleaving >100,000 kD gums to average molecular weights of 0.2-10 kDaglycan deglycosylates. That permits uptake of the smaller deglycosylatesby plants. By comparison with a variety of natural sources, branchedglycan deglycosylates from invertases showed the highest feasibility,exhibiting low cost and high potency, making them suitable forcommercial production; see Table 1.

TABLE 1 Comparisons of natural sources for branched glycandeglycosylates. Relative potencies and costs after manufacturing of thefinished products from various sources were compared. In most cases,higher potency translated to lower cost. Invertase was the source of theleast cost and highest potency glycan deglycosylates suitable forproduction. Source Potency Relative Cost Invertase 1000-1,000,000 $Botanical Gums 10-100   $$$ Shellfish 5-10   $$$$ Sawdust 1 $$$$

Terminal ligands of glycans were key to their activity, renderingidentification of this part of glycan structure critical. Suitableterminal ligands of a glycan were identified in glycopyranoses, such as,galactopyranoses, glucopyranoses, and preferably mannopyranoses; alkyl-,acyl-, and aryl-substitutions thereof; and acylglycosamines. It followedthat suitable glycans were cationic, anionic and neutral polymers;aldosyls and/or ketosyls; and branched glycans with any of the aboveterminal ligands. The molecular weight sizes typically ranged from 0.1to >500 kD, preferably between 0.2 to 10 kD, and most preferably in therange of 0.5 to 2 kD.

Glycan abbreviations are as follow:

Gal means galactopyranosyl;

Glc means glucopyranosyl;

GlcNAc means N-acetylglucosaminosyl;

GalNAc means N-acetylgalactosaminosyl;

Gly means glycopyranosyl;

Lac means lactosyl;

Ara means arabinosyl;

Man means mannopyranosyl; and

Man_(n) means poly-Man.

_(j, m, n) subscripts mean corresponding chainlengths, where m=1−24 andn=1−24, unless otherwise noted. For example, GalGlcMan_(n) meansGalactopyranosylGlucopyranosylMannopyranosyl_(n) and Glc_(m)Man_(n)means Glucopyranosyl_(m)Mannopyranosyln.

Hyphenated numerals denote the range of sizes. For example,Man₈₋₁₄GlcNAc₁₋₂ means the branchedmannopyranosyl₈₋₁₄N-acetylglucosamine₁₋₂ in which Man₈₋₁₄ means a rangeof 8 to 14 Man units in the branched chain.

Examples of preferred branched chains include Man_(n)Gly with alkyl,acyl, aryl, polyacyl, polyalkyl, amine or no substitutions; aldosyls;ketosyls; GlcNAc_(n); alkylGlc_(n); methylGlc_(n); methylGlcGly_(n);alkylMan_(n); Gal_(n)Man₂; pentoses; arabinoses; riboses; xyloses;hexoses; mannoses, mannosides, mannans; glucoses, glucosides, glucans;galactoses, galactosides, galactans, raffinoses; Gly₂, for example, Glc₂sucroses, trehaloses, maltoses, gentiobioses, cellobioses, GalGlclactoses, xylobioses, laminaribioses; Gal_(m)Man_(n);Gal_(m)Glc_(j)Man_(n); Glc_(j)Man_(n); XyloMan_(n); AraMan_(n);AraGal_(n); fructofuranosyl_(m)Glc_(j); Lac; Maltopyranosyls; Man₁₋₃,such as, Man₁, methyl-α-D-Man, methyl-α-D-Man₁₋₃, methyl-α-D-Man₃Gal;triosyls, such as, Man₃; and derivatives and combinations thereof.

Suitable glycans in a blend of deglycosylates of the glycan compositeare, for example, Man₁, Man₃, methyl-D-Man_(n) and methyl-D-Glc_(n).Glycan deglycosylates may be selected from short chains, such as,Man_(n), where n is from 1 to 8, preferably Man₁₋₃;O-linked-branched-chains, such as, Man_(m-n)Gly, Man_(m-n)Glc,Man_(m-n)Gal, and Man_(m-n)GalGlc, where m is from 1-8 and n is from1-8. Yet other suitable glycans are >1 kDa chains; such as, for example,Gal_(m)Man_(n); Gal_(n)Man_(n); Glc_(m)Man_(n); Gal_(j)Glc_(m)Man_(n),j=1−24, m=1−24, n=1−24; deglycosylates are preferred <1 kDa where j=1−8,m=1−8, n=1−8, such as for example, Gal₂Man₂; and in combinations andblends. Preferred glycan deglycosylates are O-linked glycans andN-linked glycans with Gly_(m-n) where m=1−8 and n=1−8. Further examplesinclude the following Man_(n) core structures such as α-mannobioses,mannotetraoses, mannopentaoses; amino-functionalized-mannoses, forexample, glycylMan_(n), alanylMan_(n), and aminylMan_(n), where n=1−24.

Preferred branched glycan deglycosylates may comprise one or moreN-linked-glycans such as for example Man_(n)GlcNAc₁₋₃ andMan_(m-n)GlyNAc₁₋₃, for example, Man₈₋₁₅GlcNAc₂. PreferredN-linked-glycans are selected from low molecular weightMan_(n)N-glycans, therefore, Man₃GlcNAc₁₋₃ andtrimannopyranosyl-N-glycans are most highly preferred. Suitablederivatives may have higher orders of branching, such as,Man_(m-n)GalNAc₁₋₃ for example, Man₈₋₁₅GlcNAc₁₋₂ and Man₉₋₂₀GlcNAc₁₋₃;and derivatives, such as N-glycan, acyl, alkyl, and aryl-substitutions.Furthermore, suitable branched GalGlcManN-glycans include Glc:Gal:Man inratios between 1:2:16-9:2:20; as, for example, Gal₄Man₁₀GlcNAc₂. Inaddition, N-acetylglycosaminyl-terminal ligands such as,N-acetylgalactosamines, N-acetylglucosamines, and N-acetylneuramines,may be selected from GalNAc₁₋₃; GlcNAc₁₋₃; GlcNAc₂; Man₁₋₈GlcNAc₁₋₃;derivatives and combinations, thereof.

Transition Metal²⁺ Coordination Complex Components

Embodiments disclosed herein provide transition metal²⁺ coordinationcomplex compositions of the glycan composite. In certain embodiments,the transition metal²⁺ coordination complex is comprised of the metals²⁺component and one or more anionic components. Specific metals²⁺ areincorporated into the holoprotein structure for proper binding ofglycans. In the absence of specific metals²⁺, the protein structure isincomplete, lacking the conformation to conjugate. Therefore, thesepreferred metals²⁺ include calcium (Ca²⁺) and manganese (Mn²⁺), and bothapplied together are preferred because Ca²⁺ and Mn²⁺ naturally occur inholoprotein binding sites. However, suitable transition metals²⁺ otherthan Mn²⁺ may be added, substituted or formulated including one or moreD-block transition metals²⁺ selected from cobalt (Co²⁺), nickel (Ni²⁺),and zinc (Zn²⁺); and combinations thereof; and always in the presence ofCa²⁺. In addition, the presence of iron (Fe²⁺) and magnesium (Mg²⁺)and/or one or more of the aforementioned D-block transition metals²⁺ mayfurther support the structural conformation of the holoprotein by Ca²⁺and Mn²⁺. These metals²⁺ and/or their water-soluble salts may bemeasured into the glycan composite as liquids or solids; for example,applied in the ranges of 0.1-100 ppm Ca²⁺, 0.1-100 ppm Mg²⁺, 0.1-10 ppmFe²⁺, 0.1-10 ppm Mn²⁺, 0.1-10 ppm Zn²⁺, 0.001-1 ppb Co²⁺, and 0.001-0.1ppb Ni²⁺.

Preferred anionic components of the aforementioned transition metal²⁺coordination complexes of glycan composites may be selected fromsequestering anions that further function as respiration accelerators,as follow: oxaloacetates; acetates; aconitates; citrates, isocitrates;fumarates; glutarates, ketoglutarates; malates; and succinates. Suitableacid derivatives thereof, named herein without exclusion of others, wereselected from aconitic, citric, fumaric, glutaric, malic, oxaloacetic,succinic, and like acids of transition metal²⁺ coordination complexes;and preferably at 10:1 anion:cation molar ratios or greater, within therange of 100 ppb to 30% w/w. Aconitic acids include aconitates, cis- andtrans-aconitic acids, salts, and the like. Citric acids includecitrates, citric, isocitric, and methylcitric acids; citric acidanhydrides; citric phosphates; salts, and the like. Fumaric acidsinclude fumarates, fumaric acids, boletic acids, alkylfumarates, saltsand the like. Glutaric acids include glutarates, ketoglutarates,glutaric acid, glutaric anhydride, alkylglutarates, glutamates, salts,and the like. Malic acids include malates, malic acids, maleic acids,maleates, maleic anhydrides, alkylmaleic anhydrides, maleyl-proteins,salts, and the like. Oxaloacetic acids include acetates, acetic acidssuch as glacial acetic acid and vinegar; acetyl-CoA; acetylphosphate,acetic anhydrides; alkylacetates, alkylacetoacetates, oxaloacetates,salts, and so forth. Succinic acids include succinates, succinic acids,Sprit of Amber, alkylsuccinic acids, succinyl anhydride, salts, and thelike. The aforementioned salts include, one or more of Ca, Mg, Na, andtransition metal²⁺ coordination complexes with the aforementioned acids.Anionic components of transition metal²⁺ coordination complexes may bepreferably selected from their phosphates, such as, for example,malate-phosphate, citrate-phosphate, and the like. Generally, purecomponents of transition metal²⁺ coordination complexes are commerciallyavailable in bulk. Anionic components of transition metal²⁺ coordinationcomplexes may be selected from suitable polydentate chelants, such asalkylamide chelants as follow: ammonium, sodium, and/or potassium saltsof alkyl amide chelants such as, ethylene diaminetetraacetic acids(EDTA), N-hydroxyethylethylenediaminetriacetic acids (HeEDTA),ethylenediamine-N,N′-bis2-hydroxyphenylacetic acids (EDDHA),di(ortho-hydroxybenzyl)-ethylenediaminediacetic acids (HBED),diethylenetriaminepentaacetic acids (DTPA); methylglycine N,N-diaceticacids (MGDA); glutamic acid diacetic acids (GLDA); and the like. Theanionic components are conventionally added to liquid solutions of themetals at a minimum of 7:1, and preferably at 10:1 anion:cation molarratios or greater.

Preferred salts of the transition metal²⁺ coordination complexes mayresult from reacting metal²⁺ and anion components. Moreover, suitablecommercially available salts include derivatives of N, P, K, S, C, H, O,Cl, secondary and micronutrients; and other agriculturally compliantcombinations of compounds known to the art. For example, N as amines,amides, nitrates, polyacylamines; C as carbonates; Cl as chlorides; P asphosphates, phosphites; S as sulfates; H as acids; OH as bases; and thelike.

Contributions of Exemplary Components to Functions

Glycan composites (GC) are composed of several compounds having distinctchemical characteristics, each contributing desirable properties to thewhole. It has been most elucidative to discover that when components ofthe glycan composite are applied separately to crops in the field,performance is inconsistent. Although direct application of eachcomponent to separate plant populations is possible, it is not preferredfor lack of beneficial effect. Therefore, experiments were undertaken toverify functional exemplary glycan composites; and, at the same time,showing that each of the separate components did not function adequatelyalone.

Photosynthetic crops grow by means of respiratory metabolism ofphotosynthates to build, for maintenance and to reproduce. However, theratio of respiration to photosynthesis is less than a third. Regulationof crop growth through more efficient transfer of photosynthates torespiration than before, by application of glycan composites inaccordance with embodiments disclosed herein, were used to raise thatratio. This was expedited by efficient treatments with glycan compositesin the dark (i.e., in an environment where sunlight does not reach);whether to seeds and roots underground during the day; or to rootsand/or shoots at night, for example. It would be of benefit toagriculture to enhance productivity by optimizing treatments with glycancomposites to seed, fruit, flower, sap nectar, photosynthate, root, stemand/or trunk; that is, via shoot and/or root applications, through thesenovel systems, as well. Taken together, application of glycan compositesto achieve a positive effect is realized by embodiments disclosedherein.

In contrast to photosynthetic leaves, seeds are entirely respiratory. Asa consequence, hastening germination has resulted from applications ofexceedingly low doses of GC to seeds as compared to nutrient control.Early growth responses to separated components were compared to theunified GC and results were analyzed for statistically significantcompared means. It was clearly indicated that separate components didnot work; but together, they contributed to efficacy. Furthermore, thetransition metal²⁺ coordination complexes of the embodiment thatincluded the full set of D-block transition metals²⁺ improvedperformance of composite formulations.

Respiration is dependent on available oxygen (O₂) and O₂ was enhancedthrough co-application or otherwise exposing crops under GC treatment toelevated O₂. Site-directed O₂ enhancement, in particular to roots orseeds, was achieved more efficaciously in the field by application ofO₂-generating compounds such as peroxides. Suitable inexpensiveperoxides include H₂O₂ and carbamide peroxides, while O₂-generatinggranular compounds are known to the art, such as, CaO₂ and/or MgO₂ thatslowly released O₂ while crops were under GC treatments. CaO₂ and MgO₂provided O₂ enriched environments that supported respiration,particularly when applied to seeds or roots, as separate oxygen sourcesin conjunction with formulations of the embodiment. Peroxides areO₂-generating components that may be formulated into dry products, butpreferably are stored and applied separately to the crop before, duringor after treatments with GC. Peroxides tend to destabilize and decomposeGC-concentrates, thus, shortening shelf life.

An exemplary O₂-generating co-application method follows: Prior totreatment with GC, liquid H₂O₂, 10-100 grams granular CaO₂ and/or MgO₂was incorporated at 15-30 cm soil depth at a rate of 50 Kg/ha during thecrop season; and/or potting media or planting hole soils were mixed with10-20 g/L prior to transplanting. GC was applied as a side dressing orspray drench to the photosynthetic organism to the same acreage.Thereafter, while the plant was under GC-treatment, the peroxide slowlyreleased O₂, advancing respiration in conjunction with the actions ofGC. Injecting O₂ gas may be undertaken, primarily by bubbling intoliquid media to saturation. Co-applications of O₂-generators with glycancomposites were synergistic, resulting in enhanced quantities ofproductivity.

Under circumstances in which O₂ could not be elevated, alternate methodswere applied to provide an environment conducive to respiration whereinphotosynthetic organisms under cultivation. Thus, treatments with glycancomposites were undertaken in conjunction with exposure of crops torespiration accelerators, either by addition to or incorporation withglycan composites. Respiration accelerators were selected from thefollowing: iP, such as for example, salts of phosphoric acid, such as,ammonium, potassium and sodium phosphates; Gly-phosphates such asGlc-phosphates and Man-phosphates, Glc₂-phosphates such asmannobiose-phosphates, sucrose-phosphates, trehalose-phosphates, andxylobiose-phosphates; plant growth regulators, such as, auxins; andoxaloacetic, aconitic, citric, fumaric, glutaric, malic, and succinicacids. The aforementioned acids also serve as anionic components of thecoordination complex, added at a minimum of 10:1 anion:cation ratio.

Materials and Methods

Germination and early growth of Burpee Sweet Corn cv. Bi-Licious Hybridwas tested for response to various components of composite formulations.Rapid assays of seedling growth were undertaken with hydroponic culturesin which aqueous media were sterilized and cooled. Seeds were examinedto exclude aberrantly large, small, or damaged seeds, prior totreatment. Plants were maintained in the dark for respiration at 30° C.Exactly 48 seeds were sown per 15 cm sterile disposable plastic Petridish on Whatman paper circles moistened with nutrient control ortreatments. Replicates numbered 8 per treatment (n=8). Germination wasestablished when radicle emergence was observed for 50% of controlsafter 30 hours. Treatment and control solutions were prepared bydissolving nutrients in deionized ultrapure water. In place of stainlesssteel vessels and stirrers, plastic laboratory utensils were utilized toprevent intrusion of Ni²⁺ and other metals. Cross-contamination withnutrients was avoided by disposing of plastic utensils immediately afteruse. Stock solutions were from reagent grade compounds. The aqueousglycan stock solutions of the GC in this investigation were 1-15%Man_(n) glycan deglycosylates that were obtained by acetolysis of ivorynut flour in acetic acid:acetic anhydride:sulfuric acid 25:25:1. The0.0001-5% Ca²⁺ and transition metals Fe²⁺, Mn²⁺, Zn²⁺, Co²⁺, and Ni²⁺(Cat) were prepared by sequestration in a blend of 1 mM citrate, 5 mMmalate, and 1 mM succinate, abbreviated CMS. The GC was applied at 10 μMMan_(n) concentration. Ca- and Mn-EDTA salts are limited ions,abbreviated EDTA. Other concentrations applied were 1 μM Man₃ and 100 μMMan₁. Man₁-CatCMS was formulated with Cat-CMS transition metal²⁺coordination complexes for comparison to Man₁-EDTA, and so forth. Waterwas provided as a negative control.

Results

As presented in Table 2, corn seeds treated with GC showed highlysignificant acceleration of germination mean counts (ρ=0.000) ascompared to those of separate components, Cat, glycan, CMS alone. Countsof CMS, glycan, and Cat were the same as for water; and there was nodifference between water and Cat. Whole GC showed significantenhancement as compared to G-EDTA. GC showed borderline significantimprovement when compared to glycan with CaMn-CMS, indicating that thewhole Cat improved efficacy of the GC over the contribution of limitedions. Thus, both Cat and CMS contributed to germination in the GC.Furthermore, composite formulations in which the glycan was substitutedwith Man₁ or Man₃ with Cat CMS, showed significant improvements ofgermination as compared to the Man_(i) and Man₃ formulations withlimited ions and with EDTA salts that did not enhance respiration.

Conclusions

In the whole GC, the components contributed to respiration and growth;yet, in contrast, individual components applied separately did not work.The full complement of the transition metals²⁺ in Cat, in particular inCMS transition metal²⁺ coordination complexes, significantly improvedthe product as compared to limited ion formulas. Selection of suitableanionic components of the transition metal²⁺ coordination complexes thatfacilitated respiration contributed to the GC significantly as comparedto EDTA that did not. Remarkably, the potencies of glycan and Man₃composites were orders of magnitude greater than Man₁, both showinggermination at far lower doses than Man₁. Composites were found to beapplicable toward significant improvements of Man_(i) andMan₃formulations, an unexpected outcome of the investigations. Notably,post-germination applications of GC with transition metal²⁺-alkyl amidecoordination complexes resulted in a trend toward accumulation ofphotosynthates, particularly in environments of reduced oxygen tension.For example, applications with GC-CaMnEDDHA 1 week prior to lettuce leafharvest resulted in higher Brix than whole GC and control.

TABLE 2 Effects of components of formulations on germination of cornshowed complete glycan composites function best. Statisticalsignificance of differences between mean counts of germinated seedsgrown on various components as compared to the unified glycan compositeincluded the following: Cat = non-chelated Ca²⁺, Fe²⁺, Mn²⁺, Zn²⁺, Co²⁺,and Ni²⁺; CMS = citrate malate succinate; EDTA = Ca-EDTA salts + Mn-EDTAsalts; GC = GC Cat-CMS; G-CaMnCMS = glycan and Ca²⁺-Mn²⁺-CMS; Count =germinated seeds mean count; n = 8 for all replicates; and ρ =significance. Compared treatments Count ρ GC vs Glycan 46 vs 23 0.000 GCvs CMS 46 vs 23 0.000 GC vs Cat 46 vs 23 0.000 Cat vs Water 23 vs 230.601 GC vs G-EDTA 46 vs 36 0.043 GC vs G-CaMnCMS 46 vs 38 0.054Man₁CatCMS vs Man₁EDTA 39 vs 27 0.025 Man₃CatCMS vs Man₃EDTA 45 vs 290.016

Preservatives for Liquid Concentrates

It is often advantageous to provide glycan composite products as 10ppm-30% concentrates that may be shipped in dry or liquid form whilekept under cool, dry, dark storage conditions; but inherent to organiccompounds that comprise glycan composites is that the complex wasconsumed by various and sundry microbes as well as by plant cells.Therefore, measures must be taken to preserve the compositions fromspoilage, particularly of the aqueous products. For storage, especiallyof liquid compositions, suitable preservative agents may be incorporatedto the formula to improve the stability of products. Commercialpreservative agents include biocides and germicides, such as forexample, the following: peroxides; sodium hypochlorites; bleaches;acids; bases; oxidizing agents; formaldehyde-releasing preservativessuch as 1,3-dimethylol-5,5-dimethylhydantoin, quaternium-15, bronopol,diazolidinyl urea, Na-hydroxymethylglycinate; silver; copper acetate;permanganates; dinitromorpholines; phenolics, such as,4-Chloro-3-methylphrnol and 2-phenylphenol; thiazolinones and thepreferred isothiazolinones (IT), such as, benzoisothiazolinones (BIT),methylchloroisothiazolinones and methylisothiazolinones (MIT). IT is aphytobland antimicrobial in the range of 1-800 ppm. The preservativesare recommended for formulation into liquid glycan compositeconcentrates in the range of label rates, from 1 ppm to 1%. For example,BIT in the range between 50 to 750 ppm, preferably between 100-300 ppmin liquid concentrates was safe and effective. Thus, liquid formulationof the glycan composite may be blended with any antimicrobial, yet theymust be selected from phytobland preservatives as per embodimentsdisclosed herein. In a glycan composite product present as aconcentrated product composition in the range of between 10 ppm to 30%glycan composite for dilution prior to application to a crop ofphotosynthetic organisms for the enhancement of productivity, a suitableconcentrate comprises one or more of glycan deglycosylates in the rangeof between 1 ppm to 20% and one or more transition metal²⁺ coordinationcomplexes in the range of between 1 ppm-10% and a preservative in theweight range of 50 ppm to 1%.

Exemplary Preservative Maintains Potency of Glycan Composites

The effects of preservatives on potencies of plant growth regulatorglycan composites were compared. The experiments quantified root growth.The results showed that potency was retained after storage for one monthwith preservatives. In contrast, formulations without preservative lostactivity.

Materials and Methods

Early post-germination Swiss Chard (Beta vulgaris subspecies cicla L.,cultivar “Fordhook® Giant”) root growth was tested for response toglycan composites (GC) supplemented with the preservative, BIT.Preliminary experiments of preservatives that were narrow inantimicrobial effect, not suitable for food use, or that were phytotoxicat antimicrobial doses were excluded. Roots of Swiss Chard germlingsshowed responses of improved growth in length within a week ofapplication of GC and this corresponded to weight increases. Thebranched N-linked glycan, 100 ppm Man₃GlcNAc₂ deglycosylate, wasselected to initiate blending the glycan composite with agitation intowater. The glycan composite was further formulated by stirring 5 mMmalic acid anion component in the glycan-water solution with aqueousmetal²⁺-nitrate salts; resulting in the 5 mM malate transition metal²⁺coordination complex of 1-5 ppm Ca²⁺ and transition metals 1-3 ppm Fe²⁺,0.1-0.5 ppm Mn²⁺, 0.2-1 ppm Zn²⁺, 0.01-0.1 ppb Co²⁺, and 0.001-0.01 ppbNi²⁺. The preservative, Proxel™ GXL, was selected from IT antimicrobialsand was applied to 1 ppm to 30% liquid product concentrates for storagein the range of 100-200 ppm. Formulations were stored for a month at 35°C. prior to testing. Concentrated formulations were diluted in water asneeded immediately prior to treatment of seedlings. Solutionsincorporated reagent grade compounds of other required elements.

Rapid assays of root growth were based on modified methods of theaforementioned hydroponic culture on moistened Whatman 598 Seed Culturepaper circles and in which treatments and aqueous media were notsterilized prior to application. Root mg weights were taken with acalibrated Mettler digital balance. Terminology used herein indicatesthe omission or inclusion of nutrients, as follow: GC=10 μM glycancomposite without IT, GC−IT=GC with IT; Man₁=100 μM Man₁−GC without IT,Man₁−IT=Man₁−GC with IT; MG=375 mM methyl-D-Glc₁-GC without IT, MG−IT=MGwith IT; Man₁=1 μM Man₃−GC without IT, Man₁−IT=Man₃−GC with IT.

Preservatives Retain Potencies of Stored Formulations

Post-germination Swiss chard seedlings treated with glycan compositewithout preservative showed losses of potency of as much as half that ofthe same formulation with IT. Results of means ±SE are presented inTable 3. Doubling (2×) the concentration of composite formulationswithout IT resulted in higher root weights than control, yet at 20%lower yields than 1× concentrations with IT. Formulations without apreservative lost at least half their potencies after 1 month ofstorage. The retention of original efficacies of GC formulations withpreservatives shows a distinct improvement of all products that must bestored until end-user sales and applications.

TABLE 3 Effects of composites with or without preservative IT on rootgrowth of Swiss chard seedlings are listed in order of weight yields.Formulations were assayed after 1 month of storage and those with ITshowed higher yields than formulations without IT. Mean yields withoutIT were equivalent to those of the water control. Values are measures ofroot mean milligram (mg) ± standard error (SE). Abbreviations: GC = GCwithout IT, GC-IT = GC with IT; and so forth. Man₁ = Man₁-GC; MG =methyl-D-Glc₁-GC; Man₃ = Man₃-GC. Treatment Mean ± SE (mg) GC-IT 12 ±0.3 Man₃-IT 12 ± 0.4 Man₁-IT 11 ± 0.5 MG-IT 11 ± 0.6 2X GC 10 ± 0.5 2XMan₃ 10 ± 0.6 GC  8 ± 0.4 Man₃  8 ± 0.5 Man₁  8 ± 0.6 MG  8 ± 0.6 Water 8 ± 0.5

In the example of FIG. 1, a plant cell was exposed to the solution ofglycan composite that was transported into the cell. In accordance withglycoprotein binding affinities and specificities, the glycan compositedisplaced photosynthates from storage making them available forrespiration, growth and germination. This redirected flow of energyresulted consistently in faster germination than nutrient controls,among other features.

A suitable synthesis to make a glycan composite is as follows. Incertain embodiments, the glycan composite was formulated with one ormore of the aforementioned transition metals²⁺. To make the transitionmetal²⁺ coordination complex, one or more of the appropriate anioniccomponents, such as for example, 0.1-5 mM citric, malic, succinic,and/or oxaloacetic acids were added and dissolved in water; and thensuitable ppb-ppm metal²⁺ components of the transition metals²⁺coordination complexes were stirred in to dissolve in the aqueousformulation. Thus, for example, 1-10 ppm Ca²⁺ and 0.1-1 ppm Mn²⁺, one ormore of 1-10 ppm Mg²⁺, 1-3 ppm Fe²⁺, and 0.2-1 ppm Zn²⁺, and 0.01-0.1ppb Co²⁺ and 0.001-0.01 ppb Ni²⁺ were the added metal²⁺ components.Formation of proper transition metal²⁺ coordination complexes requiresat least 1:10 and preferably 1:25 transition metal²⁺-cation:anionratios. The glycan composite unit was completed by blending in 1 μM to500 mM glycan. Formulations that were to be stored for more than a daybefore applications to plants included label quantities of a broadspectrum preservative selected from IT, BIT, MIT, hydantoin, and thelike. The method may also comprise the step of blending one or moreagricultural surfactant/emulsifiers, and/or other agriculturaladditives/adjuvants at label quantities that achieve at least criticalmicelle concentrations, in particular, for foliar applications.

Suitable surfactants and emulsifiers include anionic, cationic,nonionic, and zwitterionic detergents; for example, amine ethoxylates,alkyl phenol ethoxylates, phosphate esters, polyalkylene oxides,polyalkylene glycols, polyoxyethylene (POE) fatty acid esters, POE fattydiglycerides, POE polymers, POP polymers, PEG polymers, proteinsurfactants, sorbitan fatty acid esters, alcohol ethoxylates, sorbitanfatty acid ester ethoxylates, ethoxylated alkylamines, quaternaryamines, sorbitan ethoxylate esters, substituted polysaccharides, alkylpolyglucosides (APG), APG-citrates, alkylglycosides such asmethylglucosides, alkylmannosides, methylmannosides, ethylacetoacetates,N-acetylglucosamines, meglumines, glucamides, dimethylglucamines,copolymers, siloxanes, tallow amines, and blends. When applying glycancomposites to foliage, the formulation may further comprise one or moreaqueous surfactants and applying the resulting mixture by spraying,misting, fogging or electrostatics to the plant foliage in an amountbetween about 1 to 100 gallons per acre, preferably 10 to 80 gallons peracre.

Blending the GC with nutrients sustained vigor, expanded root systems,enhanced plant growth, enlarged floral displays, promoted fruiting andimproved flavor; a rendering that was particularly important innutrient-deficient soils and water. Essential primary elements include,N—P—K. Essential secondary nutrient elements include Ca, Mg, and S.Essential micronutrient elements include B, Cl, Co, Cu, Fe, Mn, Mo, Ni,Si, Na, and Zn. Preferred nutrients are not selected to the exclusion ofother elements, ions, or salts, and, depending on the situation, may beavailable in the soil and water in particular abundance such thatsupplementation is unnecessary for productivity; therefore, nutrientsupplementations are applied at rates compliant with governmentagricultural regulatory agencies following instructions on labels.Suitable sources include salts and minerals generally known to the art.For example, the most highly preferred micronutrient selections to thecompositions may include 0.5-5 ppm chelated Fe and/or Zn; and applying asuitable amount of the resulting mixture to one or more plants.

Plant nutrient phosphorus may be obtained from one or more of thefollowing sources: iP, phosphorus rock, phosphoric acids, phosphates,phosphites, pyrophosphates, steric P, guano, manures, seaweed extracts,bird droppings, fishery, poultry and livestock waste and the like.Organic sources of P tend to be far too expensive to apply to fields inthe 1-20% ranges of iP, however, at ranges below 1% concentration theyoften proved effective as respiration accelerators. Organic sources of Pincluded, for example, glycerophosphates and the aforementionedsugar-phosphates were utilized in the range of between 0.1 ppm to 800ppt.

Nitrogen may be obtained from one or more of the following sources:nitrate N, such as, nitric acid and salts, thereof; ammoniacal N, suchas ammonia, UAN, ammonium nitrate, ammonium sulfate; urea N such asmethylene urea, urea-formaldehyde, urea, low biuret and preferablyultralow biuret urea; amine/amide/amino N, such as alanine, arginine,asparagine, aspartate, cysteine, glutamate, glutamine, glycine,ornithine, proline, selenocysteine, taurine, tyrosine, histidine,isoleucine, leucine, lysine, methionine, phenylalanine, threonine,tryptophan, valine; salts, such as sodium or potassium glutamate;derivatives; blends; and the like; and mixtures of amino acids;proteins, such as from, glutens, caseins, fish, and blood; hexamines;and combinations, thereof. Nutrient elements were metabolized to allbiochemical, growth, reproductive and structural components ofphotosynthetic organisms.

Applications of glycan composites provide opportunities to supplementplant nutrient deficiencies by use of tank mixes or by applicationsformulated with a selection of additives. The amounts of plant nutrientswere applied in accordance with labeling of guaranteed analysis bygovernance boards at rates known to the art. Supplementation comprisedthe steps of dissolving plant nutrient components into aqueous solution,resulting in a mixture comprising glycan composites. For example, incertain embodiments, glycan composites were supplemented with 100-2000ppm N, 50-250 ppm P; 50-2000 ppm K; 1-100 ppm Ca; 1-10 ppm Fe; 0.5-6 ppmMn; and/or 0.5-5 ppm Zn. Various glycan composite formulations may beclassified as plant biostimulants that may benefit nutrient useefficiency, improve tolerance to abiotic stress, and increase yields.Biostimulants generally include natural products, such as for example,plant, algal, fermentation and animal metabolites; humates; microbials;biochemicals; and the like. Other glycan composite formulations may beclassified as elicitors that generally include natural products, such asfor example, algal, fermentation and animal metabolites; proteins;enzymes; microbials; and the like.

Embodiments disclosed herein as applied to crops are of glycancomposites made into compositions of which the compounds themselvesserve as resources for control of photosynthetic organisms, includinglive photosynthetic Eukaryotes; and, for the most part, agriculturalArchaeplastida plants. As such, the glycan composite may beappropriately formulated with agrochemicals and are rendered intoaqueous compositions that are capable of facilitating the growth ofphotosynthetic organisms, particularly cultivated flowering plants. Themethods disclosed herein may be applied to safely enhance the balancedmetabolism of exogenous components that contribute to the productivityof photosynthetic organisms. While, at the same time, they may bemethods for applications of compositions of the glycan composite in thebeneficial treatments of photosynthetic organisms, that, further, may bemade to enhance the healthy growth and quality of crops.

Although the present inventor is not to be bound by any theory, theglycan composites of the embodiments disclosed herein feature a “lockand key” mechanism. A glycan with a suitable terminal ligand shows highbinding affinity to a specific receptive glycoprotein. The lock is theglycoprotein and the key is the terminal ligand. The “tumblers” of theglycoprotein-lock are the internal structural sites of calcium andmanganese. When the glycan key opens the lock, photosynthates aredisplaced to accumulate or to move on to respiratory metabolism; andthus, opening the option for selection of respiration accelerators inthe anionic components of the transition metal²⁺ coordination complex.During daylight hours, roots consume those photosynthates. Respiringroots produce carbon dioxide, a portion (10% to 20%) of which istransported up to the shoots; and enhanced photosynthetic carbonfixation results. Thus, the glycan composite may be useful to reduceenergetic losses to photorespiration by these treatments ofphotosynthetic organisms through their reach into sap nectar thatmodulate quality of sweetness measured as Brix. In particular, when, forexample, plants are cultivated under photorespiratory environmentalstresses such as saturated sunlight intensities, droughts and heat, theglycan composite may benefit yields. Moreover, the embodiments provideglycan composite systems for regulating the accumulation ofphotosynthates through the deceleration of respiration for flavor andnutritional enhancement by human consumers, livestock, poultry, and aswell as, in robust nectar for pollinators.

In accordance with embodiments disclosed herein, a novel plant growthregulator system is introduced that advances photosynthetic flux todrive a photosynthetic organism to accumulate photosynthates. This isinitiated, for example, by creation of novel crop inputs of glycancomposites that may be applied to photosynthetic organisms. Glycancomposites may be comprised of branched glycans with transitionmetal²⁺-polydentate anions. The preferred anions of this embodiment arealkyl amides selected from salts of EDTA, EDDHA, HeEDTA, DTPA, HBED,MGDA, GLDA, and the like. In addition, oxygen-starved environments underreduced oxygen tension, in the range from 0-10% O₂, may be inducedphysically by respiration decelerators, such as by flooding roots withirrigation water, by storing plants or their parts in nitrogen gas, orby elevating CO₂ concentrations. Alternatively, respiration deceleratorsmay be selected from suitable plant growth regulators, co-appliedaccording to agriculturally labeled methods known to the art; forexample, plant growth regulators at 1-750 ppm dosages were selected fromvarious suitable cytokinins, salicylic acids, and/or, gibberellins; andderivatives and the like.

Photosynthetic organisms respond consistently to glycan compositecomponents when applied, preferably suitably formulated and renderedinto potent formulations such that they facilitate the growth andquality of photosynthetic organisms as well as provide an array ofphysiological benefits that enhance their marketable qualities.

In accordance with embodiments disclosed herein, the complexes ofembodiments may be applied separately, serially, or simultaneously.Indeed, particularly during physiological stress, by action of glycancomposites on sap nectar of a photosynthetic organism, productivity wasenhanced by the increased flow of photosynthates to respiratorymetabolism in photosynthetic organisms in accordance with theaforementioned lock and key mechanism of the embodiments disclosedherein.

In certain embodiments, concentrated glycan composite products incompositions comprising at least glycan deglycosylates in the range ofbetween 0.1 ppm to 30% and one or more transition metal²⁺ coordinationcomplexes in the range of between 0.1 ppm to 20%, are made ready forapplication by dissolving an amount of glycan composites in thepreferred carrier, water. Alternative carriers include, for example,vegetable and mineral oils, alkyl acetoacetates, or aliphatic alcohols.Therefore, it is made convenient for the grower to stir the finalsolution containing glycan composites into water as the carrier ofchoice for final dilution. In most instances, agitation and 25-80° heatfacilitates the dissolution of the dry product in the carrier. Theglycan composite is amenable to water-borne agricultural systems, suchas, hydroponic and water cultures, by metered application with pumpsinto the medium, immersion of roots in diluted glycan composites, or asa foliar treatment.

The formulations employed may include any of a wide variety ofagronomically suitable additives, adjuvants, or other agriculturallycompliant ingredients and components (hereinafter “additives”) that canimprove or at least do not hinder the beneficial effects of the glycancomposite when applied at label rates. Generally accepted additives foragricultural application are periodically listed by the United StatesEnvironmental Protection Agency. In particular, foliar compositions maycontain spreaders present in an amount sufficient to further promotewetting, emulsification, even distribution and penetration of the activesubstances. Spreaders are typically organic alkanes, alkenes orpolydimethylsiloxanes that provide a sheeting action of the treatmentacross the phylloplane. Suitable spreaders include paraffin oils and theforegoing surfactants. Penetrants include, for example, alkylacetoacetates, sodium dodecylsulfate, formamides, DMSO, and alcohols.

Embodiments herein are useful when blended or tank mixed with variousplant treatments such as agriculturally compliant pesticides,insecticides, herbicides, plant growth regulators, fungicides,germicides, biocides, elicitors, biostimulants, antagonists,antitranspirants, synergists, systemics, surfactants, spreaders,stickers, vitamins, minerals, salts, solvents, genetics, bioagents, andthe like. Herbicides that are based on ammonia metabolism, for examplethe glufosinates, Ignite®, Rely®, and Liberty®, are safened byapplication of glycan composites, reducing phytotoxicity in relatedherbicide-resistant GMO crops; and per application at label rates.

Examples of suitable additives and adjuvants include the following:minerals such as limestone, iron filings, and the like; salts such asammonium nitrate, ammonium sulfate, potassium phosphate, calciumpermanganate, calcium-phosphates, calcium acetates, calcium aconitates,calcium citrates, calcium citrate-phosphate, calcium fumarates, calciummalate, calcium malonate, calcium maleate, calcium malate-phosphate,calcium gluconates, calcium glutarates, CaO₂, calcium succinates,calcium chelants, calcium nitrate, calcium glycerophosphate,manganese-phosphates, manganese acetates, manganese citrates, manganesefumarates, manganese glutarates, rhodochrosite manganese carbonates,manganese oxides, MgO₂, manganese malate, manganese malonate, manganesemaleate, manganese succinates, manganese chelants, and the like;co-solvents such as alcohols, ketones, oils, lipids, water, and thelike; genetically modified organisms and genetic materials such as Bt,genes, sequences, RNA, DNA, plasmids, genomes, and the like; bioagentssuch as microbes, yeasts, bacteria, viruses, vectors, and the like; andcolorants, dyes, and pigments such as annatto, methylene dyes, cobaltblue, and indican. Other constituents that may be added to thecompositions include soil conditioners, antibiotics, plant growthregulators, GMO, gene therapies and the like. Among the plant growthregulators which may be added to the formulations of the presentinvention are auxins; brassinolides; cytokinins; gibberellins;salicylates; benzyladenine; amino acids; benzoates; carboxylic acids,vitamins; carbohydrates; herbicides, such as, phosphonomethylglycines,halosulfuron alkyls; selective herbicides, such as, sethoxydims andsulfonyl ureas; salts, esters, phosphates, hydrates and derivativesthereof; and genetic compositions.

Glycan composite technology is appropriate for, but not limited to, cropapplication in the dark or shade, as during periods of maximumrespiration; as well as under direct sunlight. In general, glycancomposites are readily applied directly to shoots and/or roots and/orseeds; and/or parts, thereof, including cuticle, epidermis, flower,fruit, sap, nectar, bark, stem, foliage, needle, blade, phylloplane,spine, trichome, root hair, tap root, cotyledon, cone, and the like. Theconcentration of glycan composites in the formulations as applied tophotosynthetic organisms should generally be between about 1 ppb to 1%and more preferably between about 10 ppb to 500 ppm. For specificapplications, the concentration at the point of applications may belower for roots than for shoots; thus, between the concentrations of 1ppb-300 ppm for root application. Glycan composites may be applied torooting media and then watered in or may be diluted first in an aqueouscarrier and then applied to the media. On foliage, treatments generallyare applied in a mist, fog, spray, drip, stream, dip, coating, orsprench between 1 ppb to 1% concentrations of the glycan composite. Whendiluted in an aqueous carrier, the resulting diluted glycan composite isapplied to a photosynthetic organism in an amount of about 1 to 500gallons/acre.

The following examples are provided to illustrate the embodimentsdisclosed herein and should not be construed as limiting. In theseexamples, purified water was obtained through reverse osmosis;transition metal²⁺ coordination complex components and surfactants wereobtained from Brandt. Abbreviations used in the following examples aredefined as follows: “° ” means ° C.; “Sil” meansorganosiloxane/copolymer blend; “12-26-26” means Brandt 12-26-26 Micro,N—P—K with B, Cu, Fe, Mn, Mo, and Zn; “αManda” means methyl-α-D-Man_(n),n=1-3; “GG” means combinations of branched O-linked Gal₁₋₁₂Man₂, frompartially hydrolyzed guar gum; “Ag” means GlcNAc₁₋₃; “Ethan” meansbranched Man₃GlcNAc₂ (FIG. 2); “Cat”means a blend of soluble 1 ppm Fe²⁺,0.5 ppm Mn²⁺, 0.5 ppm Zn²⁺, 0.01 ppb Co²⁺, and 0.01 ppb Ni²⁺; “CMS”means 0.1-5 mM citrate, malate, maleate and/or succinate transitionmetal²⁺ coordination complex; “IT” means isothiazolinone preservatives;“MnCO3” means manganese carbonate; “AMS” means ammonium sulfate; “MKP”means monopotassium phosphate; “DKP” means dipotassium phosphate; “MAP”means monoammonium phosphate; “DAP” means diammonium phosphate; “NH₄OH”means ammonium hydroxide; “KOH” means potassium hydroxide; “Ca(OH)₂”means calcium hydroxide; “L” means Liter; “ml” means milliliter; meansmilligram; “g” means gram; “Kg” means kilogram; means milliMolar;“Micronutrient” means trace quantities of soluble B, Ca, Co, Cu, Fe, Mg,Mn, Mo, Ni, Zn; and KOH, Ca(OH)₂, NH₄OH, MnCO₃, MAP, DAP, MKP and DKPare plant nutrients and buffering agents.

The following are examples of specific formulations that mayadvantageously be employed in methods to treat photosynthetic organismssuch as plants and to enhance growth in the same. The following examplesare intended to provide guidance to those skilled in the art and do notrepresent an exhaustive list of formulations within the scope of theembodiments disclosed.

EXAMPLE 1 Glycan Composite Formulation for Application to Roots

Ingredient Range g/L Preferred g/L Ethan 10 ppb to 1 ppm 100 ppbAconitic acid 0.001-10 0.05 Range ppm Preferred ppm Ca²⁺   1-10 5 Cat0.01-10 3

This formulation may be further supplemented with components oftransition metal²⁺ coordination complexes selected from citrates,fumarates, glutarates, malates, oxaloacetates, succinates; and Mg. Rootglycan composites were dissolved into 1 L of water with stirring at roomtemperature, 25 to 35° C.; and adjusted by titrating KOH to pH 5-7. 50to 450 gallons/acre applied as close to the roots as possible either byside dressing and/or through drip irrigation. With irrigation, thetreatment was watered into the soil, toward the roots for enhancedphotosynthates, quality and quantity.

EXAMPLE 2 Foliar Glycan Composite Formulation

Ingredient Range g/L Preferred g/L Malic acid  0.7-50 3.5 GG 0.001-10  1AMS 0.6-3 3 Sil 0.3-3 0.8 Range ppm ppm Ca²⁺    1-100 1-5 Cat 0.1-6 1-3

Ca²⁺ and Cat were dissolved with malic acid in 1 Liter of water. Otheringredients were added, dissolving each, one at a time; and the solutionwas adjusted within a range of pH 5 to pH 5.5 by titration with DKP/MAP,as needed. Transition metal²⁺ coordination complexes were selected from0.01 ppb Ni and Co; and transition metal²⁺ coordination complexes mayinclude ppm to ppt aconitates, citrates, fumarates, glutarates,oxaloacetates, and/or succinates. Foliar sprays were applied to glisten,approximating 75-100 gallons/acre resulting in enhanced photosynthates,quality and quantity of harvests.

EXAMPLE 3 Field and Flower Formulation of the Glycan Composite

Upper Preferred Range Ingredient g/L g/L 12-26-26 0.3 10 Ag 0.1 0.5 Ca²⁺0.005 1 Succinic Acid 0.7 5.0

Field and Flower Formulation Components were pre-blended as dryconcentrates to create a kit for which the dry components were storedand, later, dissolved together in water. For admixture, components wereblended in 1 L water until dissolved. The solution was applied directlyto roots. Supplementation with 0.5-3 g/L CMR or other agriculturalwetting agents was undertaken for foliar application. Field Formulationwas preferably applied as a foliar spray to shoots of plants at 50 to100 gallons/acre for enhanced photosynthates, quality and quantity. Thissolution may be further supplemented with transition metal²⁺coordination complexes selected from Ni, Co; aconitic, citric, fumaric,glutaric, malic, and oxaloacetic acids; EDTA, EDDHA, HeEDTA, DTPA, HBED,MGDA, GLDA; Mg; and the like.

EXAMPLE 4 Foliar Concentrate

Ingredient Range % Preferred αManda 1-20 5-15 Cat 1-10 5 Ca²⁺ 0.001-5   0.03 Citric acid 1-50 5-15 MKP/DAP, pH 5 1-25 5-15 BIT preservative0.01-0.8  0.1

This formulation may be further supplemented with anionic components oftransition metal²⁺ coordination complexes, such as for example,respiration accelerators selected from aconitic, fumaric, glutaric,malic, oxaloacetic, succinic acids, and the like. Alternatively, foraccumulation of photosynthates, anionic components were selected frompolydentate chelants such as, EDTA, EDDHA, HeEDTA, DTPA, HBED, MGDA,GLDA, and the like.

All components were blended to homogeneity in aqueous solution withrapid agitation until completely dissolved and adjusted to pH 5-5.5 withMKP/DAP. For foliar application, this formulation was supplemented with0.05% Sil for shoot treatments at 20-100 gallons/acre for enhancement ofphotosynthates and respiration in a crop.

An exemplary foliar system follows: Bell pepper sprouts were matched andmaintained in half-gallon plastic containers each, separated into equalpopulations of Treated and Nutrient Controls. The glycan composite fromthis example was diluted to 1% with water and applied to shoots of theTreated population as a foliar spray, while the shoots of the Controlpopulation was sprayed with the same concentrations of mineral nutrientsin water. In all other ways, Control and Treated populations werecultivated side-by-side under identical field conditions. At harvest,the Treated population averaged 35% bell pepper fruit mean weight yieldincrease over the control population that proved statisticallysignificant ρ=0.001; n=30. In addition, sun scorched peppers were absentfrom treated fruit as compared to controls that showed 1-5% loss fromscorched fruit that were not marketable due to unattractive appearance.Thus, enhanced flow of photosynthates resulted in an increase ofmarketable yields attributable to enhanced aesthetic quality of thefinal product by treatment with the glycan composite system.

The regulation of flow of photosynthates by glycan composites comprisedof aManda was further managed by optional coapplications of therespiration accelerator, 10-100 g CaO₂ to soil near roots, for highqualities and quantities of yields.

EXAMPLE 5 Exemplary Foliar PGR

Component Range % Preferred % GG 0.01-10 1 Malic acid:maleic anhydride 0.7-50 1 Water   5-80 53 KOH pH 5-7 pH 5.5 Cat-malate 0.01-1  0.1Ca-glutarate  0.01-0.3 0.1 Sil 0.3-3 0.8

Mixing Directions For malolysis, to 50% aqueous malic acid:maleicanhydride 1:1, add GG, and 1% sulfuric acid and heat to 70° C. for 8-24hours. Add the remaining dry crystals into 0.5 L of water with stirringand after completely dissolved, add the liquid solution into the aqueoussolution with rapid agitation, such as stirring. Bring the total volumeto 1 L with the addition of water. Adjust to desired pH with NH₄OH orKOH.

Mixing Directions After the components are mixed together, they arediluted in water as needed and applied as a foliar spray to shoots ofplants, preferably at between 10-100 gallons/acre. This solution may befurther supplemented with anionic components of transition metal²⁺coordination complexes selected from aconitates; citrates; fumarates;glutarates; malates; oxaloacetates; succinates; and polydentatealkylamide chelants for enhanced photosynthates, quality and quantity.

EXAMPLE 6 System for Acceleration of Germination

Ingredient Range g/L Preferred Glutaric acid  0.07-10 3 Ethan 10 ppb-1ppm 100 ppb KOH pH 5-7.5 pH 5.5 Cat 0.01-5 0.1 Ca²⁺ 0.01-1 0.05

This solution may be further supplemented with anionic components oftransition metal²⁺ coordination complexes selected from ppm-pptaconitic, citric, fumaric, malic, oxaloacetic and succinic acids;ppm-ppt polydentate alkylamide chelants; and ppm-ppt Mg.

Radish seeds, 25 per dish, were sown in 16 replicate Gosselingermination dishes on Whatman 598 Seed Culture paper circles moistenedwith Nutrient Control or glycan composite. Seeds were maintained aconstant temperature of 27° in the dark for respiration only.Germination was established at the time at which radicle emergence wasobserved for 50% of the seeds, G₅₀. Results showed accelerated glycancomposite mean G₅₀=15 hours as compared to Nutrient Control mean G₅₀=22hours; n=8; p=0.001. Treatments of radish by coating seed with 20-50μg/seed dry weight glycan composite proved highly potent, significantlyaccelerating germination as compared to Nutrient Controls as a result ofenhanced flow of photosynthates. Similar acceleration of germination wasobserved for radish seeds that were pre-coated with 20-50 μg glycancomposite/seed and dried, as compared to nutrient controls.

EXAMPLE 7 System for Enhanced Roots

Ingredient Range g/L Preferred g/L Fumaric acid 0.001-20 0.03 GG0.001-1  0.1 Cat 0.01-5 0.01 Ca²⁺ 0.01-1 0.05

This solution may be further supplemented with anionic components oftransition metal²⁺ coordination complexes selected from aconitic,citric, fumaric, malic, oxaloacetic and succinic acids; polydentatealkylamide chelants; Mg; and in ppm-ppt amounts. Bell Pepper seeds weresown in 12 replicate plots of uniform sandy loam with label rates of12-26-26. The glycan composite was formulated in water and adjusted topH 5.5 with DAP and Control was also adjusted with equivalent P withDAP/MAP. Two weeks after germination, six randomly selected plots weresprenched with glycan composite, targeting the plantings; and otherwiseall 12 plots of bell peppers received identical growth maintenance. Inorder to eliminate crowding, each plot contained 20 plants, spaced 50 cmapart. Plants were harvested with roots intact after 2 weeks; followedby clipping roots from shoots, thoroughly washing off soil andoven-drying. Root dry weights of individual plants were taken. Resultsshowed glycan composite root mean dry weight 1.2 grams as compared toNutrient Control root mean dry weight 1.0 gram; n=6; ρ=0.02. Treatmentsignificantly enhanced root dry weights as compared to controls. Sunscorched fruit, though present in controls, were absent from treatedplants indicative of enhanced photosynthates and quality of the harvest.

EXAMPLE 8 Exemplary Glycan Composite Enhancement of Hydrostatic Pressure

Fields of Plumeria flowers are conventionally cultivated under high˜1500-1700 μEinstein/m²/sec light intensity and low to moderate ˜20-30%humidity. Under these environmental conditions, daily hydrostaticpressure responses were observed in the afternoon. Typically, in theearly morning, flowering plants are high in hydrostatic pressure, but bymid-afternoon, leaves begin to droop. This cycle of midday wilt showinghigh to low hydrostatic pressures was visually distinguishable as theelevation of leaves changed from pointing upward to downward,approximately 5 to 20 millimeters (mm). An increase of hydrostaticpressure is prerequisite to growth and was measured according to thechange of elevation of foliage, especially during midday. The purpose ofthe trial was to record changes in hydrostatic pressure of Plumeria bymeasuring mm changes of elevation of leaves and comparing the responsesof aqueous nutrient controls against plants to which a single treatmentof 1-25 ml of 0.1-5 ppm Ethan glycan composite was applied to roots. Theglycan composite formula was from the aforementioned Example 6; andoptionally, other exemplary glycan composites may be applied forenhancement of hydrostatic pressure. For example, an effective amount of1-200 ml of 0.1-5 ppm partially hydrolyzed invertase deglycosylates wasapplied to roots at 8 AM and a subsequent rise in elevation of foliageshowing enhanced hydrostatic pressure was observed by noon whencontrols, at the same time, showed reduced hydrostatic pressure.Plumeria obtusa L. variety obtusa plants in 4 L plastic containers wereallowed a week to acclimate to environmental conditions of directsunlight and were further observed for consistency of diurnal changes inhydrostatic pressure with evening irrigation once per week. Mid-weekbetween watering, the baseline elevations of leaves were measured latein the morning and marked against rulers; and later, compared against mmelevations of the same leaves 5 hours after treatment.

Results: Foliar elevation was a visually discernible sign of enhancedhydrostatic pressure. The mean +15 mm rise of leaves treated with glycancomposite was significant (n=6; ρ=0.003) as compared to a correspondingmean −5 mm drop in elevation of nutrient control foliage. Aftertreatment with 10 ppb to 800 ppm glycan composites, the rise of foliagewas greater in plants treated late in the afternoon, after 3 PM; whileat the same time, control foliage showed the most pronounced loss ofhydrostatic pressure and drop in elevation caused by midday wilt.

In conclusion, Plumeria responded to treatments with increasedhydrostatic pressure when controls showed decreases. Similar increasesof quality of crop hydrostatic pressure occurred in bell pepper,brassicas, curcubits, pomes, and root crops treated with glycancomposites formulated with suitable anionic components of transitionmetal²⁺ coordination complexes. For example, anionic components wereselected from one or more of 10-900 ppm aconitic, fumaric, glutaric,malic, oxaloacetic, and succinic acids; and 10-900 ppm polydentatealkylamide chelants such as EDTA, EDDHA, HeEDTA, DTPA, HBED, MGDA, GLDA,and the like. Growth and development of all photosynthetic organisms aredependent on cell expansion initiated by increased hydrostatic pressure.When hydrostatic pressures increased over the long duration byapplication of the glycan composite systems of embodiments disclosedherein, the results showed significantly enhanced growth and developmentof the treated photosynthetic organism. Glycan composite systemscoapplied with respiration accelerators improved quantity yields ofcrops.

EXAMPLE 9 Exemplary Glycan Composite System for Low Light Intensity

Canola vegetative growth of nutrient controls in the shade was comparedto shaded populations of Canola treated with branched N-linked glycancomposite; and furthermore, treated and controlled populations werecultivated without shade to determine if there was a beneficialproductivity enhancement in relatively reduced light environments.Canola seeds were sown in 36-cell plastic flats and showed even growthafter 1 month, when shade control and glycan composite-treatments ofshaded plants were placed under 50% shade cloth. Plants that were notshaded were under natural midday full light intensity in the range of1500-1800 μEin/m2/sec; and under 50-85% shade cloth, the low lightintensity was in the range of 100 to 900 μEin/m2/sec or less than halffull light intensity.

Foliar treatments with foliar surfactants were applied, spray to drip˜100 gallons/acre. Full daylight and shaded nutrient controls werecompared against foliar glycan composite applied to Canola plants underthe same conditions. The glycan composite composition was dissolved inwater in the following order: 1-100 μM Ethan; 0.1-25 mM citrate; 0.1-2ppm Mn-CMS; 1-25 ppm Ca-CMS; and with adjustment to pH 5.5 with DAP, DKPor KOH. Plants were harvested after two weeks and dried at lowtemperature in a 70° oven for 48 hours. Dry shoots of individual plantswere weighed and mean dry weights and T-tests of equality of means ofpopulations are summarized in the table, below.

In conclusion, the population of Canola under Shade Control showedreduced productivity as compared to Full Daylight Control and Shadepopulations that were treated with glycan composites. Moreover, whenshaded plants were treated with glycan composites, benefits of theenhanced quality and quantity of productivities as compared to ShadeControl were statistically significant.

Shade Shade Glycan Fully Daylight Control composite Control Mean dryweight (g) 0.214 0.256 0.270 Increase over Shade Control 19.5% 20%Significance p = 0.000

EXAMPLE 10 Exemplary Process for Glycan Deglycosylates fromGlycoproteins

Many glycoproteins contain glycans that may be processed from seeds oflegumes, such as Jack Bean; or by deglycosylation from a number ofenzymes. Invertase is a commercially manufactured enzyme from bakersyeasts with suitable glycan components of the interior core or exteriorprotrusions that may constitute up to three quarters of the totalglycoprotein weight. Invertase has preferred branched glycans withterminal Man-ligands from such as GalManProteins; Man_(n) such asMan₁₋₆, Mannotrioses, Mannotetraoses, and Mannopentaoses;Gal_(n)Man_(n), such as, Gal₂Man and Gal₂Man₂; Gal_(n)Man_(n)-N-glycans,such as, Gal₂Man₄GlcNAc and Gal₄Man₁₀GlcNAc₂; Man_(n)-N-glycans, suchas, Man₃GlcNAc and the preferred Man₁₋₁₅GlcNAc₂; and the like. At 270kDa, the glycoprotein was too large to penetrate foliage. Therefore, toestablish a baseline for comparisons to enzymatic digestions, alaboratory process was undertaken primarily to release glycandeglycosylates from glycoproteins, preferably from invertase.

Invertase was denatured in dilute 0.2N NH₄OH, pH 12, 80° for 10 min, andneutralized by titration with 0.2N HCl; predigested with 3% trypsin at37° overnight; and further denatured by boiling for 10 minutes. Theseprotease-treated samples showed moderate 50μM germination activity inthe glycan composite. The samples were deglycosylated by incubation with200 milliunit endoglucosaminidase H at 37° for a day; and denatured indilute NH₄OH pH 12 80° for 10 min. Residual protein and peptides wereprecipitated and removed. Deglycosylates typically comprised blends ofO-linked and N-linked mannans and as glycan components of the glycancomposite glycan isolates or blends, thereof, exhibited a range of0.01-1 ppm w/w activity.

The aforementioned deglycosylations by means of proteolytic andglycolytic enzymes were relatively mild, yet involved costlybiochemicals. It was found by these experiments that the initialdenaturation of glycoproteins with base substantially shortened thesubsequent heating duration in acids, and in consideration of energysavings, is preferred. Therefore, the preferred process was byhydrolyzing glycoproteins in components that were commercially availablein bulk quantities at relatively low costs. Preferred methods weretested that may be utilized, such as, treatments with acid/base,hydrolysis, hydrazinolysis, and/or fermentations. For example,acetolysis (acetic acid:acetic anhydride:sulfuric acid 25:25:1) ofinvertase resulted in potent deglycosylates that showed activity as lowas 1-100 ppb. Optionally, a novel malolysis of the embodiment wasapplied by incubation of invertase in maleic acid:aceticanhydride:nitric acid 25:25:10 at 60-80° for 1-24 h. Alternatively,invertase was incubated in citric:phosphoric acids 25:1; saturatedcitric and/or succinic acids; and/or selected from 0.1-3N mineral acids,such as sulfuric acid, and preferably, was deglycosylated by directnitrolysis in 1-3 N nitric acid; and incubated with stirring for 1 to 24h at 40-80°.

The preferred method to partially hydrolyze invertase was as follows:10-30% invertase was dissolved in alkaline aqueous solution, such as,0.2-1 N KOH and/or 0.2 N NH₄OH with heating to 40-80° for 1-24 h; 50-60%citric acid was stirred in and incubated 1-24 h, 40-80°. Afteracid-incubation, the solution was adjusted to between a range of pH 3-6.The partially hydrolyzed invertase, now deglycosylated, was formulatedto achieve field application with at least, 1-5 ppm Ca²⁺, 0.5-1 ppm Mn²⁺and preferably with 0.01-6 ppm D-block transition metals²⁺ selected fromFe²⁺, Ni²⁺, Co²⁺, Zn²⁺; and 1-5 ppm Mg²⁺. Anionic components oftransition metal²⁺ coordination complexes were selected from one or moreof aconitates, citrates, fumarates, glutarates, oxaloacetates, andsuccinates. Alternatively, anionic components may be selected from theaforementioned polydentate chelants. For storage of solutions, one ormore preservatives were added. Enhancement of quality and quantity ofcrops of photosynthetic organisms resulted from treatment of said cropswith the aforementioned solutions containing in the range of 1 ppb to 10ppm invertase deglycosylates. For example, when a glycan compositecontaining invertase deglycosylates was applied to photosyntheticorganisms at 100 ppb partially hydrolyzed invertase the dose was 0.1mg/L.

All of the aforementioned deglycosylation methods rendered similarpotencies in their glycan composites. Elimination of any one of thecomponents of the glycan composite decreased activity. The glycancomposites comprised of invertase deglycosylates showed orders ofmagnitude higher potency than the processed botanical gums of theembodiments. Moreover, the manufacturing processes for invertasedeglycosylates were simpler and more cost effective than those of othersources. The regulation of flow of photosynthates by glycan compositescomprised of invertase deglycosylates was further instituted by optionalcoapplications of ppm respiration decelerators for flavor enhancement orrespiration accelerators for enhancement of quantity yields.

EXAMPLE 11 Exemplary Concentrate for Pollinators

Invertase was denatured in aqueous basic solution by blending 200 gramsinvertase into 1 Liter aqueous 0.2 N KOH and steamed for 4 h. Withstirring, 50% citric acid was added and the solution was heated to 80°for 12 h. After cooling, the solution was titrated to pH 6 with NH₄OH;and this was followed by addition of 1% Ca, 1% Mg, 1% Cat; and QID 2 Lwith water. The final 10% partially hydrolyzed invertase solutioncontained a blend of 3-7% branched O-linked and N-linked deglycosylatesincluding those shown in FIG. 2. This concentrate was adjusted in therange of pH 4 to 8, preferably pH 5, by addition of appropriate volumesof bases and/or acids as selected from nutrients such as KOH, NH₄OH,MnCO3, calcium carbonate, seashell flours, HCl, H2SO4, phosphoric acids,MAP, DAP, DKP, MKP, and the like; and preferably oyster shell flour.Vigorous crop growth was supported by the presence of nutrient elementsthat may be formulated at the following preferred rates: primary plantnutrients N—P—K, each 1-25%; secondary nutrients 0.1-1% Ca, 0.05-0.5%Mg, 0.1-1% S; and/or micronutrients 0.0001-0.02% B, 0.0001-0.1% Cl,0.0001 ppb-0.005% Co, 0.001-0.05% Cu, Fe 0.01-0.3%, 0.02-0.1% Mn, 0.01ppb-0.005% Mo, 0.001-0.05% Zn, 0.001-0.1% Na, and 0.0001-1 ppb Ni. Theglycan composites preferably may be supplemented with D-block transitionmetals²⁺ selected from one or more of Fe, Mn, Ni, Co, Zn; and anioniccomponents selected from respiration accelerators and polydentatechelants, and the like. In consideration of storage of liquidconcentrates, a preservative was essential to the liquid glycancomposite concentrated formulation, such as for example, selections fromaforementioned IT, in the range of 1 ppm to 800 ppm, preferably BIT inthe range of 100-200 ppm.

Not only was availability of the full body of plant nutrients importantto the crop, it was also essential to the vigorous health ofpollinators, such as honey bees, butterflies, moths, beetles, birds andbumble bees for provision of their full spectra of nutrients when theydrew nectar containing vitamins and minerals from plants. In crops ofphotosynthetic organisms, Co²⁺ is a D-block transition metal²⁺ of theembodiment that is metabolized to Vitamin B₁₂. Correction of thisdeficiency by foliar applications of glycan composites containing, forexample, Co-CMS, conferred health benefits, particularly as pollinatorsand grazers consumed photosynthetic organisms and sap nectar fortifiedwith B₁₂ and other nutrients from healthy photosynthetic crops. Forexample, when 0.0005% Co was formulated as a suitable transition metaland applied to a flowering crop, its fortified metabolites becameavailable from a flower to a honeybee and to its colony. Immediatelyprior to application to a crop, the concentrate was diluted in water orother agriculturally approved carriers in the range of 1 ppb to 100 ppm,preferably between 10 ppb to 10 ppm. The resultant diluted aqueoussolution was applied as a spray drench or root application. For foliarapplication, the product was supplemented with one or more selections ofagriculturally approved foliar surfactant and/or additives. One or moreapplications were applied per season, preferably 1-2× per month. Forfurther optimization of the flow of photosynthates to sweetness andnectar production, glycan composites were applied within a week ofblossoming and harvesting in conjunction with respiration decelerators,ppm cytokinins and ppm gibberellins, for endogenous enhancement offlavor and fortified plant foods.

EXAMPLE 12 Exemplary Invertase-based Glycan Composite (IGC) InvertaseDeglycosylate Glycans Blends of Glycan Composites

Ingredient Range % Preferred % Invertase Deglycosylates 1 ppb-100 ppm0.1-10 ppm Cat  0.01-10 0.05 Citric acids 0.001-20 0.001-15    Ca²⁺0.001-1  0.005 Ureas 0.001-25 1-15 IT preservative 0.001-0.2 0.01 Water,QID to 100%

The IGC may be further supplemented with components of transitionmetal²⁺ coordination complexes selected from D-block transition metals;anionic components; and Mg in trace quantities. Respiration acceleratorsmay be selected from, for example, one or more of aconitic, fumaric,glutaric, malic, oxaloacetic, and succinic acids. Optionally, anioniccomponents may be selected from one or more of polydentate chelants suchas EDTA, EDDHA, HeEDTA, DTPA, HBED, MGDA, GLDA, and the like.

IGC Enhances Tomato Quantity and Quality

Under stressful arid conditions for cultivation of tomato, IGC wasapplied to plants and compared against nutrient controls. IGCsignificantly enhanced general growth, yield and quality of treatedtomatoes over controls. Tomato seeds treated with IGC seed coats showedspeedy germination.

Materials and Methods

Tomato cultivar Steak Sandwich Hybrid seeds (Burpee®) seeds weregerminated under automatically controlled environmental conditions at32° in the dark. A week after germination, sprouts were transplanted outof doors for culture in an arid environment of 45° :32° LD; 10% relativehumidity; 16:8 h LD; and mostly sunny days with photosyntheticallyactive radiation (PAR) up to 1800 μmol photons m⁻² s⁻¹ at midday.Application of solutions to test plants and control plants under studywere made simultaneously and all plants were subjected to identicalconditions consistent with good laboratory practices. Chemigations weremaintained at sufficiency to keep roots uniformly moistened and drainedwithout water damage. Replicate populations of plants were cultured inplastic TLC Pro 606 trays, each of the 36 cells with 125 ml capacity,until transfer to 33 cm diameter plastic pots containing soil-lessmedia. Control and treated tomatoes were matched for size and vigor; andrunts, damaged or diseased plants and seeds were discarded prior toonset in order to produce uniform replicates. Volumes for the threefoliar applications were calibrated to 200 L/Ha with 0.1-1 ppm InvertaseDeglycosylate glycans blend content in the applied Glycan Composite.Sweetness as an indicator of the level of flavor was measured as Brix ofsap nectar squeezed from individual tomatoes using a calibrated digitalrefractometer (Reichert).

IGC was made according the methods described in Example 11 modified tothe composition in the IGC Concentrate Table, above. For foliarapplications, 0.03-0.05% Sil foliar surfactant was supplemented at labelspecifications. By application of identical overhead sprays of solution,the same quantities of nutrients were given to all plants.

Seeds were germinated on water-moistened Whatman 598 paper in Gosselindishes, 20 seeds/dish, and 5 replicates per treatment. Experimentalseeds were coated with 0.1 mg IGC, air-dried 48 h and sown. Controlswere treated with the same nutrients without IGC.

Germination was determined by radicle emergence in 50% of the seeds.After 7 days, sprouts were transplanted into flats.

Each survey pool held replicates for statistical analyses. Thedifferences between treated and control populations were statisticallysignificant in each experiment unless otherwise noted; error bars show95% confidence intervals.

Results

IGC-coated tomato seeds showed accelerated germination. All replicatesof treated seeds showed 50% germination in 60 h. In contrast, Controlsshowed 50% germination in all dishes after 72 h.

Experiments were designed to determine fruit quality and yield responsesto IGC under stressful arid conditions. Field treatments under theaforementioned conditions consistent with decelerated respiration 2 daysprior to harvest were compared to nutrient controls to determine effectson sweetness. Regardless of color, twelve of the largest >50 mm diametertomatoes were harvested 2 days after treatment and live shoot weightswere recorded. Half of the controls were red, whereas, all of thetreated tomatoes were red. Table 4 shows that appropriate treatment ofendogenous photosynthates of sap nectar resulted in enhanced fruitquality, expressed as mean Brix 5.5 that was significantly (ρ=0.012)improved as greater flavor than nutrient control Brix 4.9.

TABLE 4 Brix: Enhanced flavor quality of tomato Mean Brix Mean BrixControl IGC n = 12 4.9 5.5 p = 0.012

The count of tomatoes per plant is a measure of fruit yield and Table 5shows that IGC treatment resulted in 3.2 mean fruit count per plant thatwas significantly (ρ=0.000; n=12) greater than the mean fruit count of1.6 per nutrient control plant. The results of treatment with IGC wereimproved yield, enhanced fruit sap nectar and increased sweetness andflavor qualities in this arid environment.

TABLE 5 Average tomato fruit counts were enhanced by IGC Fruit MeanCount Fruit Mean Count Control IGC n = 12 1.6 3.2 p = 0.000

Average total fruit mean weights per tomato plant were analyzed forNutrient Control and IGC as measures of yield. Table 6, shows IGCtreatment resulted in 277 grams wet/14 grams dry weight fruit mean yieldper plant that was significantly (ρ=0.002/p=0.004) higher than control124 grams wet/14 grams dry mean fruit weight per plant.

TABLE 6 Tomato fruit weight per plant was enhanced by IGC Wet WeightMean Control IGC n = 10 124 grams/plant 277 g/plant p = 0.002 Dry WeightMean Control IGC n = 10 7 grams/plant  14 g/plant p = 0.004

Responses of various species were surveyed with results displayed inTable 7. Applications were effective on plants known for C₃ and C₄metabolism.

TABLE 7 Survey of plants that benefitted from IGC Plant Mode GeraniumRoot Petunia Sprench Lantana Root Green Tea Root Impatiens Foliar BellPepper Foliar Radish Root Coffee Root Turf Sprench Corn Seed Coat

Conclusion

Rapid germination after treatment of seeds with IGC was an indication ofimproved respiration and explained why species of CAM, C₃ and C₄responded to glycan composites because all plants respire. Endogenousmodulation of photosynthates flux by action of the glycan composite wasconsistent with tomato Brix elevation of sap nectar to improve flavorquality. Under environmental stress of the arid zone, tomato cultivationby treatment with glycan composites showed enhanced quality and quantityas compared to control. The regulation of flow of photosynthates by IGCwas further instituted by optional coapplications of the respirationdecelerator, 10-200 ppm cytokinin, for flavor enhancement or respirationaccelerators for enhancement of quantity yields. Furthermore, underconditions of O₂-starvation, supplementation with the O₂-generator,30-100 ml H₂O₂ per plant maintained root health for consistency of highqualities and quantities of yields.

EXAMPLE 13 Exemplary Partially Hydrolyzed Guar Gum (GG)

Ingredient Range % Preferred GG 1-90 3-20 Cat 1-10 5 Ca²⁺ 0.01-3    1Malic acids 0.1-50   5-25 Ureas 1-25 5-15 BIT 0.001-0.2     0.05 Water,QID to 100% QID

This formulation may be further supplemented with ppb-ppm D-blocktransition metal²⁺ coordination complex components selected from Zn, Co,Ni; aconitic, citric, fumaric, glutaric, oxaloacetic, and succinicacids; alkylamide chelants; and Mg²⁺.

Photosynthetic organisms thus treated with glycan composites comprisedof GG-deglycosylates resulted in enhanced flow of photosynthates forimprovement of the quality and quantity of harvests. The regulation offlow of photosynthates by glycan composites comprised of GG was furtherestablished by optional coapplications of respiration decelerators forflavor enhancement or respiration accelerators for enhancement ofquantity yields.

EXAMPLE 14 Exemplary Partially Hydrolyzed Tara Gum (HTG)

Tara gums are commercially available in bulk quantities and this speciescontains large polymers of branched GalMan₃ units suitable fordeglycosylation by the aforementioned acetolysis processes. That is,glycan deglycosylates were derived from food grade tara gum byacetolysis (acetic acid:acetic anhydride:sulfuric acid 25:25:1 v/v,60-80° C., 24-96 h) or nitrolysis (citric acid:acetic anhydride:nitricacid 20:20:10) yielding partially hydrolyzed branchedGalMan₃-deglycosylates. Glycans as partially Hydrolyzed Tara Gum (HTG)deglycosylates showed high potency in glycan composites between a rangeof 50-200 ppm concentrations. As a glycan component, HTG wasinvestigated showing activity only in the presence of the glycancomposite components, utilizing the methods as follow:

Cabbage was cultivated in environmentally controlled conditions asdescribed above in plastic flats with 36 plants; 125 cc/cell. Allaqueous foliar treatment solutions contained 0.5 g Sil/L, pH 6, andincluded Metals²⁺ in separate formulations of the following: Metals²⁺ 1ppm manganese sulfate, 1 ppm ferrous sulfate, and 10 ppm calcium nitratedissolved in water with 50 PPM DAP; Anionic component of the transitionmetal²⁺ coordination complex—300 ppm potassium α-ketoglutarate,abbreviated aKG; and 100 ppm HTG. Glycan composite components andMetals²⁺ were applied separately and blended together to test plantgrowth response to the components as compared to the holo-glycancomposite. The solutions of aKG, HTG, and aKG+HTG were dissolved in theaqueous Metals²⁺ solution; and, therefore, transition metal²⁺coordination complexes comprised of aKG, Ca²⁺ and Mn²⁺ were present inthe treatment solutions. Metals²⁺ served as the control.

Foliar treatments were applied in a volume of 10 ml/flat, n=36, atexpansion of the first true leaves. Shoots were harvested, dried andweighed.

Results of the trials in Table HTG, below, compared the mean dry weightsof cabbage shoots to that of Metals²⁺, the stock solution in which allof the treatments were dissolved. The aKG shoots were not significantly(n=36; ρ=0.057) different in yield than the Metals²⁺ control; HTG withMetals²⁺ showed a significant (n=36, ρ=0.001) enhancement as compared toMetals²⁺ alone. Finally, aKG+HTG+Metals²⁺ showed the most highlysignificant (n=36; ρ=0.000) enhancement of yield as compared toMetals²⁺. Furthermore, the holo-glycan composite showed highlysignificant (aKG+HTG v aKG, p=0.000; aKG+HTG v HTG, ρ=0.000)enhancements of yields as compared to all of the other treatedpopulations.

TABLE HTG Mean dry weight, Gram p Metals²⁺ 0.56 aKG 0.59 0.057 HTG 0.610.001 aKG + HTG 0.63 0.000

In conclusion, the glycan composite showed the most highly significantimprovements when formulated together as the holo-glycan compositeapplied to photosynthetic organisms, targeting photosynthates. Moreover,beneficial responses to holo-glycan composites were confirmed aftersupplementation with D-block transition metals²⁺; Ca²⁺; and Mg²⁺. Theregulation of flow of photosynthates by holo-glycan composites wasfurther instituted by optional coapplications of respirationdecelerators for flavor enhancement or respiration accelerators forenhancement of quantity yields.

EXAMPLE 15 Exemplary Partially Hydrolyzed Locust Bean Gum (PHLB)

Ingredient Range % Preferred PHLB 0.1-5   1-3  Cat 1-10 5 Ca²⁺ 0.01-3   1 CMS 0.1-50   5-25 AMS 1-25 5-15 BIT 1-750 ppm 75-100 ppm Water, QID

This PHLB formulation may be further supplemented with one or moreadditional D-block transition metals²⁺; anionic components; and/or Mg2+.Treating photosynthetic organisms with PHLB resulted in enhanced flow ofphotosynthates for improvement of the quality and quantity of harvests.The regulation of flow of photosynthates by glycan composites comprisedof PHLB was further controlled by optional coapplications of therespiration decelerator, 10-200 ppm salicylic acid, for flavorenhancement.

EXAMPLE 16 Methods and Compositions for Speedy Germination

Ingredient Range g/L Preferred Ethan 1 ppb-1 ppm 10-100 ppb Glutaricacid 0.0007-1   0.003 KOH pH 5-7.5 pH 5.5 Cat 0.01-5 0.1  Ca²⁺ 0.01-10.05

For germination, this formulation may be further supplemented withanionic components of transition metal²⁺ coordination complexes that arerespiration accelerators selected from one or more of aconitic, fumaric,glutaric, malic, oxaloacetic, and succinic acids; and phosphates,thereof.

Radish seeds, 25 per dish, were sown in 16 replicate Gosselingermination dishes on Whatman 598 Seed Culture paper circles moistenedwith Nutrient Control or glycan composite. Seeds were maintained at aconstant temperature of 27° C. in the dark for respiration only.Germination was established at the time at which radicle emergence wasobserved for 50% of the seeds, G₅₀. Results showed accelerated glycancomposite mean G₅₀=15 hours as compared to Nutrient Control mean G₅₀=22hours; n=8; ρ=0.001. Treatments of radish by coating seed with 20-50μg/seed dry weight glycan composite proved highly potent andsignificantly hastened germination as compared to Nutrient Controls as aresult of enhanced flow of photosynthates. Similarly, germination washastened in radish seeds that were pre-coated and dried with 20-50 μgglycan composite/seed, as compared to Nutrient Controls.

EXAMPLE 17 N-linked Glycan Composites

Ingredient Range ppm Preferred ppm Ethan 0.001 to 1 0.1 Citric acid0.001-5 0.05 Ca²⁺    1-10 5 Cat  0.01-10 3

This formulation may be further supplemented with anionic components oftransition metal²⁺ coordination complexes. For example, the anioniccomponents may be selected from respiration accelerators and polydentatechelants.

Root glycan composites were made in 1 L of water with stirring at roomtemperature, 25 to 35° C. The formulation was titrated with KOH and/orNH₄OH to pH 5-7. 5; and 500 to 750 gallons/acre were applied as close tothe roots as possible either by drench, spray-drench, sidedressingand/or through chemigation. With irrigation, treatments were wateredinto the soil toward the roots for enhanced quality and quantity.Applications were applied weekly to monthly as needed during the growingseason.

Example 18 Invertase Liquid Concentrate

Invertase glycoproteins are comprised of proteins with core and surfaceMan_(n) polymers. As such, these glycoproteins were partially hydrolyzedto yield Man_(n) deglycosylates. When formulated in glycan compositesand applied to photosynthetic organisms, invertase deglycosylatesmodulated the flow of energy from photosynthesis to respiration.Concentrated glycan composite formulations containing N-linked branchedchain deglycosylates from invertase were diluted with water to 10 ppb-5ppm field doses for photosynthetic organisms and were applied togermination assays, showing strongly potent activity at levels as low as10 ppb invertase deglycosylates.

Glycan composites were further formulated for regulation of flow ofphotosynthates with optional selections for flavor enhancement fromrespiration decelerators and enhancement of quantity yields byrespiration accelerators. Transition metal²⁺ coordination complexes ofinvertase deglycosylates may be further selected from anionic componentsthat are respiration accelerators such as from ppm aconitic, fumaric,glutaric, malic, oxaloacetic, and succinic acids. Optionally, anioniccomponents may be selected from polydentate chelants such as ppm EDTA,EDDHA, HeEDTA, DTPA, HBED, MGDA, GLDA, and the like.

Enhanced quality and quantities of crops yields resulted from glycancomposite treatments of, for example, blueberry, leafy vegetables,cotton, cereals, tomato, cherry, onion, coffee, banana, citrus, melon,leafy greens, nuts, pomes, berries, tree fruit, food crops, florals,trees, turf and ornamental crops.

Methods

To 40 ml of strong base, preferably selected from 0.2 N KOH, NH₄OH, andCa(OH)₂, 10-12 g dry invertase powder with preferred activity 200,000Sumner Units/g was added with agitation and the aqueous alkalinesolution was steamed to 60-80° C. for 2-24 hours. Next, 20 g citric acidwas added with stirring and the solution was steamed at 60-80° C. for3-12 hours. The lower the heating temperature, the longer theincubation; and this process was followed by cooling to roomtemperature. Suitable base, preferably selected from KOH, NH₄OH, andCa(OH)₂ was added with agitation to adjust to pH 4-6. Volume was broughtup to QID 100 ml to make 10% invertase deglycosylates. 1-6 ppm Cat stocksolution was prepared and 1 ml 10% invertase deglycosylates was dilutedin 1 L Cat stock solution for 100 ppm invertase deglycosylates liquidconcentrate. 100 ppm BIT was added with agitation to complete invertasedeglycosylates liquid concentrate.

Invertase deglycosylates liquid concentrate field dilutions: 10 mlinvertase deglycosylates 100 ppm liquid concentrate was diluted in 1 Lwater to yield 1 ppm invertase deglycosylates, followed by serialdilutions to 1, 10, and 100 ppb, as needed. Foliar applications requiredaddition of one or more agricultural surfactants, such as, 0.5 g/L Sil.Applications of low concentrations, in the range of between 10 ppb-5 ppminvertase deglycosylates were effective and showed significantly earliergermination and enhancement of the qualities and quantities of crops ofphotosynthetic organisms as compared to control. The regulation of flowof photosynthates by glycan composites comprised of invertasedeglycosylates was further adapted by coapplication of the respirationdecelerator, 1-100 ppm gibberellin, for flavor enhancement. Furthermore,under conditions of O₂-starvation, supplementation of 1 L media with 10g CaO₂ maintained high qualities and quantities of yields.

EXAMPLE 19 Exemplary Partially Hydrolyzed Botanical Gums

Botanical gums were selected for branched chain high mannan contentsthat were partially hydrolyzed according to the following methods: Guar,konjac, locust bean, tara and ivory nut gums were separately dissolvedin 60-80° water with agitation as 1% w/w gums. To each gum solution, 1-3N nitric acid and 5-25% acetic anhydride were added. The solutions weremaintained at 70-80° for 1-8 h. The solutions were cooled to 30-40° andtitrated to pH 3.5-4 with Ca(OH)₂, KOH and/or NH4OH. To the solutions,30-50% citric acid was added to saturation and heated to 60-80° C. for1-24 h. Solutions were allowed to cool to room temperature and titratedto pH 5-6 with Ca(OH)₂, KOH, MnCO3 and/or NH₄OH. 10× Cat was dissolvedinto the partially hydrolyzed gum solution to make 0.1% gum. Serialdilutions with Cat provided samples of various doses and were comparedwith the nutrient control solution.

Exemplary components for glycan composites comprised of partiallyhydrolyzed botanical gums are given in Table of Components, below.

Table of Components Component Range % Preferred % Botanical Gum 0.01-30.1 HNO₃:Acetic Anydride:CMS 5:5:5-25:25:60 10:10:50 NH₄OH, KOH, Ca(OH)₂pH 3-6 pH 5.5 Transition metal²⁺ 0.01-1 0.1 coordination complexCa-coordination complex 0.01-3 0.1

Bioassay

Radish seeds, 30 per dish, were sown in 20 replicate Gosselingermination dishes on moistened Whatman 598 Seed Culture paper circles.Seeds were maintained at a constant temperature of 27° in the dark tomaintain only respiration. Germination was established at the time atwhich radicle emergence was observed for 50% of the seeds, G₅₀.

Results showed faster glycan composite mean G₅₀ at 15 hours as comparedto Nutrient Control mean G₅₀ at 22 hours; n=10; p=0.001. Potencies ofvarious partially hydrolyzed (PH) gums showed general correspondence ofconcentrations necessary for G₅₀ activity to their contents of branchedmannans. Potencies were observed from low activity in PH gums of guar,konjac, locust bean, tara, and to high activity by PH ivory nut, asdetailed in the Table of Active Doses (ppm) of Partially Hydrolyzed (PH)Gums.

Treatments of radish seed with glycan composites from different sourcesof gums showed different levels efficacy and significantly hastenedgermination as compared to Nutrient Controls. Speedy germination was aresult of endogenously enhanced flow of photosynthates. Preferredrespiration accelerators included citric and malic acids. Furthermore,under conditions of O₂-starvation, root supplementation of each 1 Lmedia with 20 g CaO₂ maintained root vigor for high qualities andquantities of yields.

Table of Active Doses (ppm) of Partially Hydrolyzed (PH) Gums 100 ppm GG30 ppm PH Konjac 20 ppm PH Locust Bean 20 ppm PH Tara 10 ppm PH IvoryNut

Example 20 Exemplary Invertase Glycan Composite Liquid Concentrate

Materials: Maxlnvert 200 invertase powder (DSM)

Citric Acid, anhydrous (Brandt)

A highly concentrated liquid solution of Invertase Glycan Composite wasprepared by denaturing and deglycosylating active invertase with heatingto 80° in saturated citrate solution. A saturated 50% citric acid wasfirst prepared in water. To 500 ml saturated citric acid solution, 100 gMaxlnvert 200 invertase dry powder was slowly added into the agitated(300-800 rpm) acid solution to avoid clumping. After the invertase wasdispensed, stirring was reduced to 100 rpm to minimize foaming. Thesolution was heated to 80° ; and maintained at 75-85° for hours withslow 100 rpm stirring that resulted in deglycosylates, a blend of Glycancomponents of the Glycan Composite, from this process that partiallyhydrolyzed invertase. After cooling to room temperature, thepreservative, 100 ppm BIT was added with stirring and the total volumewas brought to 500 ml with water to make a 20% Glycan solution.

The Metal⁺² concentrate stock was prepared, containing the following:ultra low biuret urea; Ca; Mn; Fe; Zn; Mg; random block copolymeremulsifier, such as, Pluronic L-62; lower aliphatic alcohols,preferably, propanols; preservatives, such as, BIT; concentrations ofeach component per Table Invertase Glycan Composite Concentrate. Ameasure of 5 ml 20% Glycan was diluted per 10 L Metal⁺² concentratestock for 100 ppm Glycan Composite;

and titrated with citric acid, KOH, and/or NH₄OH between pH 5-8. This100 ppm Glycan Composite product was prepared for field dilutions to0.01-1% in water for applications to photosynthetic organisms. Forexample, 10 ml Glycan Composite in 1 L water yielded 1 ppm Glycan; andsimilarly, 1 gallon/100 gallons per acre or 3 fluid ounce/1000 foot².Serial dilutions also were applied, as needed. The field-diluted GlycanComposite from invertase deglycosylate was effective in a range of 0.01to 10 ppm Glycan on roots; and/or with label rates of foliar surfactantadditives, 0.1 to 10 ppm Glycan proved effective when applied to shoots.

Table Invertase Glycan Composite Concentrate pH 7-8 Input ComponentsPreferred g/10 L % Range Water 8617 60-90  20% Glycan 5 0.01-10   Urea700 5-25 Mg 2.5%, EDTA 200 1-10 Mn 6%, EDTA 50 0.3-5   Fe 7%, EDTA 1000-10 Ca 3%, Citrate 68 0.5-5   Zn 9%, Acetate 50 0.3-5   Pluronic L-62100 1-6  Isopropanol 100 0.5-10   BIT 6 0.08-0.3 

Although specific features of the embodiments disclosed herein aredescribed with respect to one example and not others, this is forconvenience only as some feature of one described example may becombined with one or more of the other examples in accordance with themethods and formulations disclosed herein.

Other permutations of the methods and formulations of the embodimentsdisclosed herein will occur to those skilled in the art and are withinthe following claims:

What is claimed is:
 1. A glycan composite comprising a branched glycandeglycosylate and one or more transition metal²⁺ coordination complexes.2. The glycan composite of claim 1, wherein said branched glycandeglycosylate has a glycopyranose or acylglycosamine terminal ligand. 3.The glycan composite of claim 1, wherein said branched glycandeglycosylate is a Man₁₋₈-Gly₁₋₈.
 4. The glycan composite of claim 1,wherein said one or more transition metal²⁺ coordination complexescomprises (a) one or more anionic components and (b) one or more metal²⁺components.
 5. The glycan composite of claim 1, further comprising oneor more preservatives.
 6. The glycan composite of claim 5, wherein saidone or more preservatives are selected from the group consisting ofbenzoisothiazolinones, methylchloroisothiazolinones,methylisothiazolinones, and combinations thereof.
 7. The glycancomposite of claim 1, wherein said branched glycan deglycosylate isselected from the group consisting of O-linked glycans and N-linkedglycans with Gly_(m-n); where m=1-8 and n=1-8; and wherein said one ormore transition metal²⁺ coordination complexes comprises both metals²⁺components and one or more anionic components.
 8. The glycan compositeof claim 1, wherein said branched glycan deglycosylate is atrimannopyranosyl-N-glycan.